Biodegradable fabric softener premix compositions are described containing (1) biodegradable softener actives that contain substantial unsaturation and (2) perfume. The premixes can be added at ambient temperatures to water containing acid, to provide a pH of from about 1.5 to about 5, to form finished compositions. The water can also contain calcium and/or magnesium salt to modify the viscosity.

Fabric softening compositions containing high solvent levels are known in the art. However, there is a need for highly concentrated "compositions" that are, in effect, premixes, that can be used to form finished compositions that are suitable for sale to consumers, said highly concentrated compositions having a viscosity that allows them to be stored, pumped, and processed at ambient temperatures into finished aqueous compositions without the need for high shear mixing. This is of particular importance when one wants to provide the fabric softening benefit to consumers in remote areas of the world, where the cost of transporting finished product to the area from some other location is prohibitively expensive and the local manufacturing facilities are limited and rudimentary.
The present invention provides highly concentrated compositions containing biodegradable actives with either no, or relatively low, organic solvent level (i.e., below about 40%, by weight of the composition), that have improved stability (i.e., do not precipitate, gel, thicken, or solidify) at normal, i.e., room temperatures and sub-normal temperatures under prolonged storage conditions, that can be processed at room temperatures, and that will disperse in water, preferably without high shear mixing, to form stable compositions.

Concentrated, stable, premix for forming fabric softening composition


The present invention allows for rendering, with a micropolygon renderer, motion blurred images of objects modeled and animated with the most commonly used class of implicit surfaces. The invention introduces the technique of automatic implicit differentiation to automatically compute the time derivatives of points on the combined surface from the time derivatives of the constituent surface elements while evaluating the combined field function that defines the composite surface.

The present invention relates generally to the field of computer graphics and computer generated animation. More specifically, the invention relates to rendering photorealistic images of moving objects modeled with implicit surfaces.
Surfaces of objects to be rendered in computer graphics can be numerically described by different mathematical models. Some surface models, e.g., NURBS (non-uniform rational B-Splines), are most naturally specified in terms of a local parameterization, i.e., a mapping from R2 to R3 in which the parameters, conventionally taken as u and v, serve as local coordinates on the surface and the location in three dimensional object space of a point on the surface is specified as an explicit function of the u and v parameters: x(p)=ƒ1(u, v); y(p)=ƒ2(u, v); z(p)=ƒ3(u, v).
Parametric surface definitions are handy but cumbersome and become more cumbersome when one attempts to model objects of non-trivial topology. To model surfaces which are not topologically equivalent to a sheet, cylinder or torus (or one of its twisted variants, e.g., a Klein bottle), one must explicitly stitch together different parameterization patches. Insuring continuity and smoothness across patch boundaries can be difficult, particularly when animating the models.
An alternative class of mathematical models that have become popular for more efficiently and elegantly modeling organic looking objects with smoothly blended curved surfaces are those defined by implicit functions, i.e., as equipotentials of a scalar field. See, e.g., J. Bloomenthal, et al., Introduction to Implicit Surfaces (1997). Complicated surfaces can be modeled by combining scalar fields that define simpler constituent surface elements. Depending on how one combines the constituent scalar fields, one can obtain surfaces that smoothly blend the constituent shapes, or are the precise union, intersection, or subtraction of the constituents or any combination thereof.
The ability to specify complicated smoothly blended shapes merely by combining simple functions is a key advantage to modeling with implicit surfaces. One is thereby able to both intuitively and efficiently build elegant and complicated surfaces. Inplicit surfaces offer additional advantages in computer generated animation. With implicit surfaces, one can model the motion of a complicated blended surface merely by specifying the motion of its individual constituent objects. In addition, one can easily model surfaces of changing topology. Depending on the distance between constituent objects and the manner in which the field functions are combined, objects will automatically combine and separate. This is particularly useful in modeling the motion of fluids, e.g., splattering raindrops.
The advantage of implicit surface animation, that one can determine the shape of the surface at anytime by merely performing the appropriate combination of field functions of the constituent elements, has the corresponding disadvantage, that one cannot relate a particular point on a surface defined at time t1 to a point on the surface at a different time, t2. Even if the motion of points on the constituent surfaces is specified (which in some cases is more information than is needed to model the surface which requires only the constituent field functions as a function of time), it is far from obvious how to determine the motion of points on the combined surface. The problem is most apparent when one considers the areas of xe2x80x9cblendxe2x80x9d between constituent surface elements, i.e., where the points being considered are on none of the consistent surfaces.
The inability to determine the motion of specific points on the surface can be a serious (fatal) hindrance to doing computer animation with implicit surfaces with certain important and widely used rendering systems. In particular, to incorporate motion blur in a micropolygon render, e.g., the REYES architecture as implemented in Pixar""s Photo-Realistic Renderman(copyright), one most be able to calculate the change in position of micropolygon vertices during the time in which the fictional camera shutter is open.
A solution to the problem of determining the motion of particular identified points on a surface defined by the equipotentials of an arbitrary computable combination of scalar fields is therefore required in order to fully utilize implicit surface modeling techniques in high quality computer animation.
The present invention allows for rendering, with a nicropolygon renderer, motion blurred images of objects modeled and animated with the most commonly used class of implicit surfaces (i.e., those formed as a combination of constituent elements for which one is able to specify the motion). The invention introduces the technique of automatic implicit differentiation to automatically compute the time derivatives of points on the combined surface from the time derivatives of the constituent surface elements (which must be supplied by the animator) while evaluating the combined field function that defines the composite surface.


The present invention is directed to a process for the production of a peptide, polypeptide, or protein having a predetermined property. In accordance with one embodiment, the process begins by producing by way of synthetic polynucleotide coupling, stochastically generated polynucleotide sequences. A library of expression vectors containing such stochastically generated polynucleotide sequences is formed. Next, host cells containing the vectors are cultured so as to produce peptides, polypeptides, or proteins encoded by the stochastically generated polynucleotide sequences. Screening or selection is carried out on such host cells to identify a peptide, polypeptide, or protein produced by the host cells which has the predetermined property. The stochastically generated polynucleotide sequence which encodes the identified peptide, polypeptide, or protein is then isolated and used to produce the peptide, polypeptide, or protein having the predetermined property.

Priority of Switzerland patent application serial no. 1379/85-8, filed Mar. 30, 1985, is claimed through PCT International Application No. PCT/CH85/00099, filed Jun. 17, 1985, under 35 U.S.C. xc2xa7xc2xa7120/365.
The present invention has as its object a process for obtaining DNA, RNA, peptides, polypeptides, or proteins, through use of transformed host cells containing genes capable of expressing these RNAs, peptides, polypeptides, or proteins; that is to say, by utilization of recombinant DNA technique.
The invention aims in particular at the production of stochastic genes or fragments of stochastic genes in a fashion to permit obtaining simultaneously, after transcription and translation of these genes, a very large number (on the order of at least 10,000) of completely new proteins, in the presence of host cells (bacterial or eukaryotic) containing these genes respectively capable of expressing these proteins, and to carry out thereafter a selection or screen among the said clones, in order to determine which of them produce proteins with desired properties, for example, structural, enzymatic, catalytic, antigenic, pharmacologic, or properties of liganding, and more generally, chemical, biochemical, biological, etc. properties.
The invention also has as its aim procedures to obtain, sequences of DNA or RNA with utilizable-properties notably chemical, biochemical, or biological properties.
It is clear, therefore, that the invention is open to a very large number of applications in very many areas of science, industry, and medicine.
The process for production of peptides or polypeptides according to the invention is characterized in that one produces simultaneously, in the same medium, genes which are at least partially composed of synthetic stochastic polynucleotides, that one introduces the genes thus obtained into host cells, that one cultivates simultaneously the independent clones of the transformed host cells containing these genes in such a manner so as to clone the stochastic genes and to obtain the production of the proteins expressed by each of these stochastic genes, that one carries out selection and/or screening of the clones of transformed host cells in a manner to identify those clones producing peptides or polypeptides having at least one desired activity, that one thereafter isolates the clones thus identified and that one cultivates them to produce at least one peptide or polypeptide having the said property.
In a first mode of carrying out this process, stochastic genes are produced by stochastic copolymerization of the four kinds of deoxyphosphonucleotides, A, C, G and T from the two ends of an initially linearized expression vector, followed by formation of cohesive ends in such a fashion as to form a stochastic first strand of DNA constituted by a molecule of expression vector possessing two stochastic sequences whose 3xe2x80x2 ends are complementary, followed by the synthesis of the second strand of the stochastic DNA.
In a second mode of carrying out this process, stochastic genes are produced by copolymerization of oligonucleotides without cohesive ends, in a manner to form fragments of stochastic DNA, followed by ligation of these fragments to a previously linearized expression vector.
The expression vector can be a plasmid, notably a bacterial plasmid. Excellent results have been obtained using the plasmid pUC8 as the expression vector.
The expression vector can also be viral DNA or a hybrid of plasmid and viral DNA.
The host cells can be prokaryotic cells such as HB 101 and C 600, or eukaryotic cells.
When utilizing the procedure according to the second mode mentioned above, it is possible to utilize oligonucleotides which for a group of palindromic octamers.
Particularly good results are obtained by utilizing the following group of palindromic octamers:
5xe2x80x2 GGAAT7CC 3xe2x80x2
5xe2x80x2 GGTCGACC 3xe2x80x2
5xe2x80x2 CAAGCTTG 3xe2x80x2
5xe2x80x2 CCATATGG 3xe2x80x2
5xe2x80x2 CATCGATG 3xe2x80x2
It is also possible to use oligonucleotides which form a group of palindromic heptamers.
Very good results are obtained utilizing the following group of palindromic heptamers:
5xe2x80x2 XTCGCGA 3xe2x80x2
5xe2x80x2 XCTGCAG 3xe2x80x2
5xe2x80x2 RGGTACC 3xe2x80x2
where X=A, G, C, or T, and R=A or T
According to a method to utilize these procedures which is particularly advantageous, one isolates and purifies the transforming DNA of the plasmids from a culture of independent clones of the transformed host cells obtained by following the procedures above, then the purified DNA is cut by at least one restriction enzyme corresponding to specific enzymatic cutting site present in the palindromic octamers or heptamers but absent from the expression vector which was utilized; this cutting is followed by inactivation of the restriction enzyme, then one simultaneously treats the ensemble of linearized stochastic DNA fragments thus obtained with T4 DNA ligase, in such a manner to create a new ensemble of DNA containing new stochastic sequences, this new ensemble can therefore contain a number of stochastic genes larger than the number of genes in the initial ensemble. One then utilizes this new ensemble of transforming DNA to transform the host cells and clone these genes, and finally utilizes screening and/or selection and isolates the new clones of transformed host cells and finally these are cultivated to produce at least one peptide or polypeptide, for example, a new protein.
The property serving as the criterion for selection of the clones of host cells can be the capacity of the peptides or polypeptides, produced by a given clone, to catalyze a given chemical reaction.
For instance, for the production of several peptides and/or polypeptides, the said property can be the capacity to catalyze a sequence of reactions leading from an initial group of chemical compounds to at least one target compound.
With the aim of producing an ensemble constituted by a plurality of peptides and polypeptides which are reflexively autocatalytic, the said property can be the capacity to catalyze the synthesis of the same ensemble from amino acids and/or oligopeptides in an appropriate milieu.
The said property can also be the capacity to modify selectively the biological or chemical properties of a given compound, for example, the capacity to selectively modify the catalytic activity of a polypeptide.
The said property can also be the capacity to stimulate, inhibit, or modify at least one biological function of at least one biologically active compound, chosen, for example, among the hormones, neurotransmitters, adhesion factors, growth factors, and specific regulators of DNA replication and/or transcription and/or translation of RNA.
The said property can equally be the capacity of the peptide or polypeptide to bind to a given ligand.
The invention also has as its object the use of the peptide or polypeptide obtained by the process specified above, for the detection and/or the titration of a liquid.
According to a particularly advantageous mode of carrying out the invention, the criterion for selection of the clones of transformed host cells is the capacity of these peptides or polypeptides to simulate or modify the effects of a biologically active molecule, for example, a protein, and screening and/or selection for clones of transformed host cells producing at least one peptide or polypeptide having this property, is carried out by preparing antibodies against the active molecule, then utilizing these antibodies after their purification, to identify the clones containing this peptide or polypeptide, then by cultivating the clones thus identified, separating and purifying the peptide or polypeptide produced by these clones, and finally by submitting the peptide or polypeptide to an in vitro assay to verify that it has the capacity to simulate or modify the effects of the said molecule.
According to another mode of carrying out the process according to the invention, the property serving as the criterion of selection is that of having at least one epitope similar to one of the epitopes of a given antigen.
The invention carries over to obtaining polypeptides by the process specified above and utilizable as chemotherapeutically active substances.
In particular, in the case where the said antigen is EGF, the invention permits obtaining polypeptides usable for chemotherapeutic treatment of epitheliomas.
According to a variant of the procedure, one identifies and isolates the clones of transformed host cells producing peptides or polypeptides having the property desired, by affinity chromatography against antibodies corresponding to a protein expressed by the natural part of the DNA hybrid.
For example, in the case where the natural part of the hybrid DNA contains a gene expressing xcex2-galactosidase, one can advantageously identify and isolates the said clones of transformed host cells by affinity chromatography against anti-xcex2-galactosidase antibodies. After expression and purification of hybrid peptides or polypeptides, one can separate and isolate their novel parts.
The invention also applies to a use of the process specified above for the preparation of a vaccine; the application is characterized by the fact that antibodies against the pathogenic agent are isolated, for example, antibodies formed after injection of the pathogenic agent in the body of an animal capable of forming antibodies against this agent, and these antibodies are used to identify the clones producing at least one protein having at least one epitope similar to one of the epitopes of the pathogenic agent, the transformed host cell corresponding to these clones are cultured to produce these proteins, this protein is isolated and purified from the clones of cells, then this protein is used for the production of a vaccine against the pathogenic agent.
For example in order to prepare an anti-HVB vaccine, one can extract and purify at least one capsid protein of the HVB virus, inject this protein into an animal capable of forming antibodies against this protein having at least one epitope similar to one of the epitopes of the HVB virus, then cultivate the clones of transformed host cells corresponding to these clones in a manner to produce this protein, isolate and purify the protein from culture of these clones of cells and utilize the protein for the production of an anti-HVB vaccine.
According to an advantageous mode of carrying out the process according to the invention, the host cells consist in bacteria such as Escherichia coli whose genome contains neither the natural gene expressing xcex2-galactosidase, nor the EBG gene, that is to say, Zxe2x88x92, EBG-E. coli. The transformed cells are cultured in the presence of X gal and the indicator 1PTG in the medium, and cells positive for xcex2-galactosidase functions are detected; thereafter, the transforming DNA is transplanted into an appropriate clone of host cells for large scale culture to produce at least one peptide or polypeptide. The property serving as the criterion for selection of the transformed host cells can also be the capacity of the polypeptides or proteins produced by the culture of these clones to bind to a given compound.
This compound can be in particular chosen advantageously among peptides, polypeptides, and proteins, notably among proteins regulating the transcription activity of DNA.
On the other hand, the said compound can also be chosen among DNA and RNA sequences.
The invention has also as its object those proteins which are obtained in the case where the property serving as criterion of selection of the clones of transformed host cells consist in the capacity of these proteins to bind to regulatory proteins controlling transcription activity of the DNA, or else to DNA and RNA sequences.
The invention has, in addition, as an object, the use of a protein which is obtained in the first particular case above mentioned, as a cis-regulatory sequence controlling replication or transcription of a neighboring DNA sequence.
On the other hand, the aim of the invention also includes utilization of proteins obtained in the second case mentioned to modify the properties of transcription or replication of a sequence of DNA, in a cell containing the sequence of DNA, and expressing this protein.
The invention has as its object as well as a process of production of DNA, characterized by simultaneous production in the same medium, of genes at least partially composed of stochastic synthetic polynucleotides, in that the genes thus obtained are introduced into host cells to produce an ensemble of transformed host cells, in that screening and/or selection on this ensemble is carried out to identify those host cells containing in their genome stochastic sequences of DNA having at least one desired property, and finally, in that the DNA from the clones of host cells thus identified is isolated.
The invention further has as its object a procedure to produce RNA, characterized by simultaneous production in the same medium, of genes at least partially composed of stochastic synthetic polynucleotides, in that the genes thus obtained are introduced into host cells to produce an ensemble of transformed host cells, in that the host cells so produced are cultivated simultaneously, and screening and/or selection of this ensemble is carried out in a manner to identify those host cells containing stochastic sequences of RNA having at least one desired property, and in that the RNA is isolated from the host cells thus identified. The said property can be the capacity to bind a given compound, which might be, for example, a peptide or polypeptide or protein, or also the capacity to catalyze a given chemical reaction, or the capacity to be a transfer RNA.
Now the process according to the invention will be described in more details, as well as some of its applications, with reference to non-limitative embodiments.
First we shall describe particularly useful procedures to carry out the synthesis of stochastic genes, and the introduction of those genes in bacteria to produce clones of transformed bacteria.

Process for obtaining DNA, RNA, peptides, polypeptides, or protein, by recombinant DNA technique


A method and system for reallocating the data blocks of a logical data entity are described. According to one aspect of the invention, the physical arrangement of data blocks of a logical data entity are analyzed to determine a fragmentation level. Next the fragmentation level is compared with a fragmentation threshold previously assigned to the logical data entity. If the fragmentation level exceeds the fragmentation threshold, only those data blocks of the logical data entity that are within a predefined allocation area having a predetermined number of contiguous data blocks that are not associated with the logical data entity are reallocated.

In a typical modern file system, when a file is initially written to a disk or other mass storage device, the file is divided into equally-sized, logical data blocks, and an attempt is made to write the logical data blocks sequentially and contiguously to the disk or mass storage device. Consequently, when the file is read from the disk or mass storage device, the individual data blocks can be read sequentially, thereby limiting the number of read operations required, which in turn limits the seek time required by the reading component (e.g., disk drive head) to search for and locate the individual data blocks comprising the file. However, over time, as existing files are modified and deleted, and new files are written, the availability of physical contiguous space for writing new files becomes limited. Consequently, files tend to become fragmented. When the files of a file system become fragmented, multiple read operations are required to read the files, and the overall performance of the system suffers.
For a variety of reasons, fragmentation issues may arise with network-based storage systems that implement write anywhere file systems, such as the WAFL® (Write Anywhere File Layout) file system developed by Network Appliance of Sunnyvale, Calif.). Storage systems having file systems with write anywhere capabilities differ from storage systems with more traditional file systems in several respects. First, the write allocation policy of a write anywhere file system is extremely flexible when compared to many traditional file systems. Consequently, the data and/or metadata that make up a particular logical data container (e.g., a file, volume, or LUN) may be written to any location on any mass storage device. For example, when a storage system with a write anywhere file system is writing a file's individual logical data blocks to a disk, the storage system may write the data blocks to any location (e.g., disk blocks) on any disk connected to the storage system. Moreover, when a particular data block of an existing file is modified, the modified data block will not necessarily be written back to the same physical location (e.g., disk block) as the initial data block.
Consequently, with a write anywhere file system, certain classes of workloads are likely to endure performance degradation due to fragmentation issues. For instance, performance levels associated with workloads that are characterized by a large number of random writes followed by a sequential read tend to suffer from fragmentation issues. This may occur, for example, with workloads associated with certain database applications, such as Microsoft® Exchange Server™. For example, during a work day, a large file that makes up an individual user's email mailbox may be modified several times, causing the large file to become severely fragmented. At night, when a verification or backup process is performed on the mailbox file, an attempt is made to sequentially read the file. However, due to the fragmentation caused by the user's activity throughout the day, the system may have a difficult time sequentially reading the mailbox file. Accordingly, reading the fragmented mailbox file is accomplished via a greater number of read operations than would be necessary if the mailbox file was not fragmented.
With a storage system having a traditional file system, a defragmentation application is often utilized to improve the overall performance when the storage system is suffering from a badly fragmented file system. A typical defragmentation application analyzes a fragmented file system to identify files that are fragmented. After identifying a particular file that is fragmented, the defragmentation application attempts to read all of the data blocks comprising the particular file, and to re-write the data blocks to contiguous unallocated disk blocks of the disk or other mass storage device. Consequently, data blocks are physically rearranged. However, the defragmentation operation does not increase the number of allocated disk blocks. For example, for every new disk block allocated, a previously allocated disk block is unallocated and made available.
Traditional defragmentation applications may be ineffective when used with certain network-based storage systems that implement write anywhere file systems. In particular, the WAFL® file system has certain characteristics, which render traditional defragmentation tools less useful. One such characteristic is the ability to create "Snapshots™" when used in conjunction with tools such as SnapDrive™ or SnapManager® for Microsoft® Exchange, both made by Network Appliance, Inc. A Snapshot™ is a form of read-only, persistent, point-in-time image (PPI) of a particular data set, such as a volume, file system, or logical unit number (LUN). The term "Snapshot" is used herein without derogation of any trademark rights of Network Appliance, Inc.
In WAFL®, a file is structured as a tree (hierarchy) of blocks, where the upper level blocks contain meta-data pointing to the lower level blocks that store the actual data. During a Snapshot operation, rather than creating an entirely new tree of blocks to form the Snapshot, only the upper level blocks containing the meta-data are copied, and these copied upper level blocks point to the original lower-level (actual data) blocks. As data in the active file system is modified, the meta-data for the active file system is changed to point to new data blocks, whereas the Snapshot does not change.
FIG. 1 illustrates an example of what occurs during a Snapshot operation. The block labeled "AF" represents a meta-data block (e.g., an inode), for a file that is part of the active file system. The "AF" meta-data block contains meta-data including pointers to four data blocks, "A", "B", "C" and "D", which store the actual data that make up the file. When originally written to disk, the data blocks are written sequentially and contiguously. During a Snapshot operation, the "AF" meta-data block is copied, and a new meta-data block is created, for example, Snapshot meta-data block "SN1". The Snapshot meta-data block "SN1" includes pointers to the same data blocks (e.g., blocks "A" through "F"), because at that particular point in time (e.g., time=2), the file that is part of the active file system is the same as the file that was captured in the Snapshot operation.
At some later point in time (e.g., time=3), when the file is modified, rather than overwriting the disk block storing data block "C" with a new data block, a new disk block is allocated (e.g., the disk block storing data block "C*"). Accordingly, the pointer in the "AF" meta-data block is updated to point to the new data block "C*". However, because the data blocks that make up the Snapshot are read-only, the pointer in the Snapshot meta-data block "SN1" continues to point to the original data block "C". Consequently, the data blocks that make up the file in the active file system (e.g., the bold data blocks) become fragmented. However, because data blocks "SN1", "A", "B", "C" and "D" are part of the read-only Snapshot, those data blocks cannot be moved or rewritten. Therefore, to completely reorder the data blocks that make up the file in the active file system, data blocks "A", "B", and "D" need to be copied to a new location, while data block "C*" needs to be relocated. However, this would result in the addition of three new data blocks being allocated. Consequently, a traditional defragmentation application, which might move the data blocks of the active file system to a new location, would be ineffective because it would nearly double the number of allocated blocks.
Furthermore, in an environment with one or more network-based storage systems, there are often multiple file systems involved. For example, each client computer system configured to utilize a storage server may have its own local file system. For example, the local file system on the client may logically map a file's data to logical data blocks, e.g., file block numbers (FBNs). In addition, the storage server may have its own file system that manages the physical mapping of the data to physical storage units (e.g., disk blocks) on the mass storage device. The client's logical mapping of the data blocks will not necessarily coincide with the storage server's physical mapping of the data blocks. Consequently, a defragmentation application executed locally on the client may improve the logical mapping of the data blocks. However, by improving the logical arrangement, the locally executed defragmentation application may cause greater fragmentation of the data blocks as physically stored on the mass storage device. Consequently a defragmentation application executed locally on the client may actually make matters worse in certain instances.

Method and system for reallocating data in a file system


The present invention relates to novel catalysts for removing heteroatoms, particularly nitrogen, from hydrocarbonaceous feedstocks. The catalysts are comprised of highly dispersed molybdenum sulfide promoted with a noble metal such that the noble metal is in an oxidation state greater than 0 and coordinated to S. The noble metal is selected from Pt, Pd, Rh, and Ir. It is preferred that the catalysts of be prepared from a precursor composition selected from platinum ethoxyethyl xanthate or platinum dithiocarbamate. Additionally, the catalyst may include a promotor sulfide such as nickel sulfide, cobalt sulfide or iron sulfide, etc. or mixtures thereof.

Hydrotreating of petroleum feedstocks and various boiling fractions thereof has become increasingly important because of more stringent product quality requirements. Furthermore, the petroleum industry foresees the time when it will have to turn to relatively high boiling feeds derived from such materials as coal, tar sands, oil-shale, and heavy crudes. Feeds derived from such materials generally contain significantly more deleterious components, such as sulfur, nitrogen, oxygen, halides, and metals. Consequently, such feeds require a considerable amount of upgrading in order to reduce the content of such components, thereby making them more suitable for further processing, such as fluid catalytic cracking and/or cracking and/or catalytic reforming.
Hydrotreating is well known in the art and usually requires treating a hydrocarbonaceous feed with hydrogen in the presence of a catalyst to effect conversion of at least a portion of the feed to lower boiling products, usually with removal of deleterious components. See for example U.S. Pat. No. 2,914,462 which discloses the use of molybdenum sulfide for hydrodesulfurizing gas oil and U.S. Pat. No. 3,148,135 which discloses the use of molybdenum sulfide for hydrorefining sulfur and nitrogen-containing hydrocarbon oils. Further, U.S. Pat. No. 2,715,603 discloses the use of molybdenum sulfide as a catalyst for the hydrogenation of heavy oils, and U.S. Pat. No. 3,074,783 discloses the use of molybdenum sulfides for producing sulfur-free hydrogen and carbon dioxide, wherein the molybdenum sulfide converts carbonyl sulfide to hydrogen sulfide. Molybdenum and tungsten sulfides have other uses as catalysts, including hydrogenation, methanation, water gas shift, etc. reactions.
In general, with molybdenum and other transition metal sulfide catalysts, as well as with other types of catalysts, greater catalyst surface areas generally result in more active catalysts than similar catalysts with lower surface areas. Thus, those skilled in the art are constantly trying to achieve catalysts having ever greater surface areas. More recently, it has been disclosed in U.S. Pat. Nos. 4,243,553 and 4,243,554 that molybdenum sulfide catalysts of relatively high surface area may be obtained by thermally decomposing selected thiomolybdate salts at temperatures ranging from about 3000.degree. to 8000.degree. C. in the presence of essentially inert, oxygen-free atmospheres. Suitable atmospheres are disclosed as consisting of argon, a vacuum, nitrogen, and hydrogen. In U.S. Pat. No. 4,243,554, an ammonium thiomolybdate salt is decomposed at a rate in excess of 15.degree. C. per minute, whereas in U.S. Pat. 4,243,553, a substituted ammonium thiomolybdate salt is thermally decomposed at a substantially slower heating rate of about 0.5.degree. to 20.degree. C. per minute. The processes disclosed in these patents are claimed to produce molybdenum disulfide catalysts having superior properties for water gas shift and methanation reactions as well as for catalyzed hydrogenation and hydrotreating reactions.
Hydrotreating catalysts comprising molybdenum sulfide, in combination with other metal sulfides, are also known. For example, U.S. Pat. No. 2,891,003 discloses an iron-chromium composition for desulfurizing olefinic gasoline fractions. Further, U.S. Pat. No. 3,116,234 discloses Cr-Mo and also Mo with Fe and/or Cr, and/or Ni for hydrodesulfurization. Also, U.S. Pat. No. 3,265,615 discloses Cr-Mo for hydrodenitrogenation and hydrodesulfurization.
Hydrotreating catalysts containing platinum are also known. For example, U.S. Pat. No. 3,422,002 discloses hydrotreating with a catalyst consisting essentially of 0.05 to 5 wt. % of a platinum series metal and about 4 to 30 wt. % of molybdena on alumina, the catalyst having been presulfided.
While various of these catalysts have met with commercial success, there still exists a need in the art for catalysts having ever improved properties with respect to hydrodenitrogenation over those conventionally used.

Multimetallic sulfide catalyst containing noble metals for hydrodenitrogenation


Methods and compositions are provided for the persistent modification of cell membranes with exogenous proteins so as to alter the function of the cell to achieve effects similar to those of gene therapy, without the introduction of exogenous DNA. DNA sequences, the proteins and polypeptides embodying these sequences are disclosed for modulating the immune system. The modulations include down-regulation, up-regulation and apoptosis.

It has long been the goal of experimental biology and medicine to induce cells to behave in predictable ways and to alter the behavior of cells in ways that are beneficial to a subject. For example, if undesired cells could be induced to alter their behavior to undergo apoptosis while normal cells retain normal function, subjects with a disease caused by proliferation of undesired cells would obtain relief from the disease. Similarly, if tissue-rejecting cells can be eliminated or their behavior changed, transplantation with tissues foreign to the subject can be successful.
Gene therapy has been proposed for selected diseases in order to correct or modify pathological or physiological processes. In gene therapy as it is generally termed, specific DNA is introduced into a tissue and organ, where it is produces various proteins that will correct or ameliorate the condition. It is an unpredictable therapy, depending on potentially dangerous expression vectors and the uncertain efficiency of delivery, which is often low. Moreover, gene therapy is considered to have dangerous side effects, such as sustained expression in desired cells or tissues past the desired duration of therapy, or the introduction of genetic modifications in undesired tissues or cells. Because of such adverse effects, as discovered in human clinical trials, much caution is advised before gene therapy is put in practice.
DNA-based gene therapy has been the subject of intense studies during the past few decades because of the tremendous potential it offers for the treatment of inherited diseases and other pathologic conditions for which the expression of selected proteins may offer treatment. For example, gene therapy can be used for immunomodulation either to enhance the capacity of the immune response to deal with infections or tumors, or to down-regulate the immune response for the prevention of autoimmunity or foreign graft rejection. Despite vast efforts in the past two decades, the safe and effective application of gene therapy to the treatment of diseases has been extremely limited. Among the apparent drawbacks of gene therapy are the possibility of causing permanent change in the DNA complement of the host; uncertain tissue specificity; expression of the encoded protein beyond the intended duration of therapy; and high cost. It would therefore be extremely beneficial if cells and tissues could be modified to express proteins of interest in a short period of time without the introduction of foreign DNA.
Whereas gene therapy has generally focused on the problem of delivering nucleic acids into cells, much fundamental knowledge concerning the important role of cell surface molecules has been gained through studies of signal transduction in cells of the immune response, which are readily accessible and have well-understood functions. The immune response is regulated by the interaction of several different cell types, which react to the presence of foreign antigens. The adaptive immune response is critical to the survival of vertebrates in an environment full of pathogenic microorganisms. Individuals who, due to inborn genetic defects, exposure to chemotherapeutic agents or infection by such viruses as human immunodeficiency virus (HIV), lack a fully functional immune response are susceptible to infections that an individual with a healthy immune response would readily withstand. However, the immune system does not always function in ways that are beneficial to the organism. Its dysregulation leads to autoimmunity and tumors. The immune system also serves as a barrier to the transplantation of foreign grafts, such as cells taken from an individual other than the transplant recipient. Transplantation permits the replacement of failed cells, tissues, or organs in otherwise terminal diseases, while bone marrow transplantation can treat hematopoietic disorders, malignancies, autoimmune disorders, and other diseases. For transplantation to be successful, it is necessary either to suppress the adaptive immunity or to "teach" the recipient's immune system to accept these foreign antigens as native.
The immune response to foreign antigens is initiated by naive T cells that use clonally-expressed T cell receptors (TCRs) to recognize antigens such as peptides presented by self-major histocompatibility complex (MIC) molecules. This recognition reaction, when accompanied by costimulatory signals provided by antigen-presenting cells (APCs), has been thought to result in full T cell activation. A productive T-cell response is now seen as requiring three distinct signals. Signal 1 is generated by T-cell receptor interaction with the major histocompatibility complex (MHC) antigen/peptide on antigen-presenting cells (APCs). Signal 2 is mediated by the engagements of costimulatory molecules, such as B7/CD28 and CD40/CD40L, on T cells and APCs. Signal 3 is transduced via cytokines elaborated by T cells and APCs that have received both Signal 1 and 2. The transduction of these 3 signals drives T cells and APCs to proliferation and differentiation into effectors for the generation of a productive immune response. The lack of any of these signals during the T-cell response results in T-cell anergy and immune nonresponsiveness. For example, tumors evade the immune system by preventing the transduction of one of these signals.
Upon activation, T cells proliferate and differentiate into effector cells that evoke immunological mechanisms responsible for the clearance of antigens from the system. A period of death then follows during which most of the activated T cells undergo apoptosis-mediated "activation-induced cell death" (AICD) and effector activity subsides. Apoptosis is a complex process that involves a series of extra- and intracellular signals that converge on the activation of enzymes called caspases that commit the cell to apoptosis.
Transplantation of foreign cells (such as bone marrow and stem cells), tissues (such as pancreatic islets), and organs (such as kidneys, hearts, livers) has become an important and effective therapeutic alternative for patients with selected terminal diseases. The transplantation of foreign grafts between genetically different patients (allografts between members of the same species or xenografts between members of different species) is, however, limited by the ability to control the immunological recognition and rejection of the graft by the recipient.
Bone marrow (BM) transplantation has been viewed as an extraordinarily promising treatment for hematopoietic and autoimmune disorders and for certain cancers. One obstacle to bone marrow transplantation is the possibility of rejection of the transplanted tissue, mediated by the host's T cells and NK cells. Graft-versus-host-disease (GvHD) is another possible adverse consequence of bone marrow transplantation. Donor T cells in the transplanted tissue can mount an immune response against the host's vital organs, often leading to death of the host. Host-versus-graft reactions and GvHD therefore limit the clinical use of bone marrow transplantation, which might otherwise be widely used to treat various diseases and to prevent foreign graft rejection.
Pharmacological agents that cause immunosuppression are now a mainstay of regimens for the control of allograft rejection. Although such drugs are effective in reducing the severity of rejection episodes, they are nonspecific and fail to create a state of permanent graft-specific tolerance. Continuous exposure of the recipient to these immunosuppressive agents is therefore associated with a significantly increased risk of opportunistic infections and malignancies. The need remains to develop more selective and long-lasting methods to prevent BM rejection.
Additionally, these nonspecific immunosuppressive agents can induce serious and undesirable side effects in the host. These adverse effects often outweigh the benefits for patients with diseases in which the body identifies certain parts of itself as "foreign" and launches an adaptive immune attack that results in autoimmunity, such as is observed in Type I diabetes, arthritis, lupus, and multiple sclerosis. It would be very desirable to be able to "teach" the immune system to tolerate the "foreign" self-antigen.
It would also be very desirable to be able to "teach" the immune system to rid the organism of tumor cells. T cell-mediated cellular immunity is the most critical acquired response against tumors. A series of experimental studies has provided evidence that tumors evade T-cell-mediated immunity by several different mechanisms. These mechanisms include: i) lack of Signal 1, due to inefficient display of MHC/tumor antigen bimolecular complexes on tumor cells or defects in the transduction of this signal; ii) absence of Signal 2, due to the absence of costimulatory molecules on tumor cells; iii) induction of anergy in T cells; and iv) physical elimination of effector T cells via apoptosis. Although all of these mechanisms may be operative in patients with a large tumor burden, the lack of costimulation is believed to play the most critical role.
The need therefore remains to develop more rapid, selective and long-lasting methods to modulate cell function without the introduction of nucleic acids into cells for therapeutic purposes. The need also remains to develop a means of accomplishing the end of gene therapy without many of the risks attendant to the introduction of exogenous nucleic acids into an organism. Since much cell function is controlled through the transduction of signals at the cell surface, a generally applicable method of attaching an agent to a surface would be useful. Among the uses of such a method would be: the modulation of cell function without the introduction of nucleic acids into the cell; the accomplishment of the end of gene therapy by alternative and potentially preferable means; and the manipulation of an organism's immune response in order to diminish that response, as to treat autoimmunity or to forestall graft-versus-host disease, or to increase that response, as to treat tumors or infections.

Alteration of cell membrane for new functions using IL-2 and streptavidin

UNIVERSITY OF LOUISVILLE RESEARCH FOUNDATION, INC.


The invention relates to a compound of the general formula (I),

Chemokines are small molecular weight (8-12 kDa) secreted polypeptides playing important regulatory role in the immune processes due to their leukocyte attracting (chemotactic) effect. They exert their effects through the chemokine receptors, which belong to the family of the G protein coupled receptors.
The CC chemokine receptors 3 (CCR3 receptors) are expressed by a number of inflammatory cells, like the basofils, mast cells, T lymphocytes, epithelial cells, dendritic cells, but in the greatest amount they can be found on the surface of the eosinofiles.
The CCR3 receptor ligands belong to the family of the C—C kemokines. They have a number of selective and non-selective ligands. The selective ligands are the eotaxin, eotaxin-2 and the lately discovered eotaxin-3. The non-selective ligands are the RANTES, the monocyte chemotactic proteins (MCP-2, MCP-3, MCP-4) and the macrophag inhibitor protein (MIP-1). The best characterized CCR3 ligand known from a long time is the eotaxin.
The eotaxin through the activation of the CCR3 receptors attracts selectively the eosinofils. Prior to an allergen provocation, the measured eotaxin level in the broncho-alveolar lavage fluidum of asthmatic patients was by 67 percent higher. On the effect of provocation a 2.4-fold increase of the epithelial and endothelial cells of the respiratory tract were found.
In the lung the eotaxin is produced in many cells. Following an allergen response, the most important eotaxin sources are the epithelial cells, but a great amount of eotaxin is produced by the fibroblasts of the lung, the smooth muscle cells and the endothelial cells of the respiratory tract, the alveolar macrophags and lymphocytes, and the eosinofils themselves.
Originally the data showed that the CCR3 receptors are to be found only in the eosinofil cells (Bertrand C P, Ponath P D., Expert Opin Investig Drugs. 2000 January; 9(1):43-52.), but on the basis of expression profiles it has been revealed that other inflammatory cells—although in smaller amount—also contain CCR3 receptors (Elsner J, Escher S E, Forssmann U., Allergy. 2004 December; 59(12):1243-58.). Thus, the CCR3 antagonists possess much wider effect, their activity is not limited to the eosinofils and consequently they can be considered much more valuable and effective targets in the treatment of asthmatic, allergic and inflammatory diseases.
Based on the above observations, CCR3 antagonists may possess important profilactic and therapeutic effects in the treatment of pathologies where in the development of the disease CCR3 receptors play a role. These diseases are characterized by the disorder of the leucocyte functions (activation, chemotaxis), there are numerous chronic inflammatory diseases among them, such as asthma, allergic rhinitis, atopic dermatitis, eczema, inflammatory bowel disease, ulcerative colitis, allergic conjunctivitis, arthritis, multiple sclerosis, Crohn's disease, HIV-infection and diseases in conjunction with AIDS.
The CCR3 antagonists published to date in the literature are carbamide-, thiocarbamide derivatives (WO 01/09088, WO 02/059081) and/or compounds containing saturated cyclic amino group (WO 00/35451, U.S. Pat. No. 6,605,623, WO 01/98270, WO 03/004487, WO 03/018556, WO 2004/028530, WO 00/53600, WO 00/35876, WO 01/64216, WO 02/50064, WO 02/102775, GB 2373186, WO 03/082291, WO 2004/004731, WO 2004/058702, WO 2004/085423). The present invention relates to a new structural type of compounds, to the open-chain amino-alkyl-amide derivatives, representatives of these compounds are effective CCR3 receptor antagonists.
From the aspect of therapeutic use it is essential that the molecules do not bind, or bind only in case of very high concentration to other the CCR receptor subtypes.
Our aim was to prepare compounds of high antagonistic activity, and at the same time selective to the CCR3 receptor, i.e. which inhibit the CCR3 receptor in much smaller concentration as compared to other CCR receptors. Further aim was that the new compounds have stability, bioavailability, therapeutic index and toxicity values which ensure its drugability. Additional aim was that the compounds, through their good enteric absorption can be applied orally.

Amino-alkyl amide derivatives as CCR3 receptor ligands

Abstract: as defined herein which is useful for the treatment of a pathology in a patient wherein a CCR3 receptor plays a role in the development of the pathology, and pharmaceutical preparations containing such compound. The invention is also directed to a process for preparing the compound of the general formula (I), and intermediate useful in the preparation.


The softness of tissue products, such as facial and bath tissue, is improved by the addition of a carboxylic acid ester. The carboxylic acid ester can be incorporated into the tissue by addition of the ester to the furnish prior to the formation of the tissue, or it can be topically applied to the tissue web after formation, either while the tissue web is still wet or after the tissue web has been dried.

In the manufacture of tissue products, such as facial and bath tissues, continual efforts have been made to make the tissues softer and more appealing to the consumer. To this end, chemical addition has been utilized more frequently as the softening effects of a variety of chemicals are discovered. Nevertheless, there remains a need for further improving the feel qualities of such tissues.

Tissue products containing esters of polyoxyethylene alkyl ether carboxylic acids


A method for manufacturing a foraminous abrasive article whereby abrasive particles are concentrated in an external surface region of a foraminous substrate. The method includes the steps of: providing a foamable, hardenable, liquid resin composition and a foraminous substrate having an external surface; foaming the resin composition effective to disperse a plurality of gas bubbles throughout the resin composition; applying the foamed resin composition to the external surface of the substrate to form a foam coating layer having an upper surface; applying a plurality of abrasive particles to the upper surface of the foam coating layer; heating the foamed coating layer effective to eliminate the gas bubbles from the foamed coating layer; and hardening the resin composition to attach the abrasive particles to the substrate. The invention also concerns the foraminous abrasive article products of this method.

Foraminous abrasive articles have been made, for example, as nonwoven abrasive articles constituted of a network of synthetic fibers or filaments which provide surfaces upon which abrasive particles are adhesively attached.
Nonwoven abrasive articles, in particular, are useful in various converted forms, such as wheels, sheets, discs, flap brushes, and the like. In these converted forms, the resulting nonwoven abrasive articles are useful to clean, condition, and/or decorate the surfaces such materials as metal, wood, plastics, glass, ceramics, composites, and the like. A particularly important use for such nonwoven abrasive articles is to scuff automotive body finishes prior to the application of further coatings.
Conventional nonwoven abrasive articles generally involve a mat of fibers which have on at least a portion of their surface an abrasive coating comprising abrasive particles and a binder. As known, the fibers can be formed from various synthetic polymers, including polyamides, polyesters, polypropylene, polyethylene, and various copolymers thereof. Also, naturally occurring fibers such as cotton, wool, bast fibers, and various animal hairs may also be suitable. Suitable abrasive particles can be formed of flint, garnet, aluminum oxide, diamond, silicon carbide, and the like. Binders commonly comprise cured versions of hide glue or varnish, or one or more resins such as phenolic, urea-formaldehyde, melamine-formaldehyde, urethane, epoxy, and acrylic resins. Phenolic resins include those of the phenol-aldehyde type.
In one conventional general scheme, nonwoven abrasive articles have been manufactured by applying to a nonwoven web starting material a "prebond" coating of binder precursor solution, which includes one or more of the above-named resins, in order to impart sufficient strength to the nonwoven web starting material so that it can better tolerate subsequent processing. A "make" coating optionally has been applied to the prebonded nonwoven web and left non-fully cured up until the time when abrasive particles are later applied to the web to help attach the abrasive grains throughout the lofty fibrous mat. Finally, an abrasive coating of resinous binder material and abrasive particles has been applied onto the nonwoven to increase the abrasive characteristics of the nonwoven. The resin binder used in the various coatings may be the same or different.
In a conventional approach, the binder coating for the abrasive particles has been applied to the nonwoven web as a non-foamed dispersion of a binder resin and abrasive particles in a liquid medium. The dispersion is then applied to the nonwoven substrate by means, such as spraying, that either atomizes the mixture to atomized droplets, or otherwise causes a film of the dispersion to be formed on the web. The atomized droplets or film are applied to the nonwoven web and cured. During curing, the droplets either flow together (coalesce) by heat-induced viscosity reduction, or cure as individual droplets where they stand. Films usually flow together to cover (wet) most of the filaments in the nonwoven web, and they are cured in position.
As the binder resin, phenolic resins are used extensively to manufacture nonwoven abrasive articles because of their thermal properties, availability, low cost, and ease of handling. The monomers currently used in greatest volume to produce phenolic resins are phenol and formaldehyde. Other important phenolic starting materials are the alkyl-substituted phenols, including cresols, xylenols, p-tert-butylphenol, p-phenylphenol, and nonylphenol. Diphenols, e.g., resorcinol (1,3-benzenediol) and bisphenol-A (bis-A or 2,2-bis(4-hydroxyphenyl)propane), are employed in smaller quantities for applications requiring special properties.
There are two basic types of phenolic resins: resole and novolak phenolic resins. Molecular weight advancement and curing of resole phenolic resins are catalyzed by alkaline catalysts. The molar ratio of aldehyde to phenolic is greater than or equal to 1.0, typically between 1.0 and 3.0. In the production of adhesive coatings for nonwoven abrasives, one standard starting phenolic resin composition is a 70% solids condensate of a 1.96:1.0 formaldehyde:phenol mixture with 2% potassium hydroxide catalyst added based on the weight of phenol. The phenolic resin composition is typically 25-28% water and 3-5% propylene glycol ether, which are required to reduce the viscosity of the resin. In conventional techniques for making nonwoven abrasives, the phenolic resin has not been applied to the nonwoven web substrate in foamed condition.
It is possible for nonwoven abrasive articles resulting from these conventional coating methods to have an abrasive particle distribution through the thickness of the web such that abrasive particles somewhat more concentrated near the external surface regions of the mat (due to spray coating). However, substantial quantities of abrasive particles nonetheless are usually present throughout the web including the interior regions. The abrasive particles lodged in the central interior regions of the web are not immediately available for useful abrading work until the external surface(s) of the article is worn or otherwise attrited.
For applications in automotive body finishing, the ability of nonwoven abrasive articles to quickly create a uniformly scratched surface is of primary consideration. For this purpose, it would be advantageous to have the abrasive particles concentrated at the external surfaces of such nonwoven articles thereby increasing the number of such particles in immediate, simultaneous contact with the workpiece, instead of being more or less uniformly distributed throughout the thickness of the web.
The state of the art can be further understood by reference to the following references in particular:
U.S. Pat. No. 2,958,593 (Hoover et al.) describes a low density, open, nonwoven fibrous abrasive article formed of fibers formed into a nonwoven web, abrasive particles, and a curable binder. Organic fibers are adhesively bonded together at crossing and contacting points, and abrasive particles are also adhesively bonded to the web fibers. The interstices between the fibers are left open and unfilled by adhesive or abrasive particles so that the web is non-clogging and non-filling in nature, and it consequently can be readily cleaned upon flushing. The adhesive used to bond the contacting points of the fibers in the web can also be used as the means of attaching the abrasive particles to fibers in the web. The abrasive mineral particles are sprayed onto the nonwoven web as dispersed in the liquid binder solution. Alternatively, the mineral binder can be roll coated, dip coated, or separately applied relative to the abrasive particles, upon the nonwoven web. For instance, the mineral binder can be first sprayed upon the web followed by sifting of the abrasive mineral particulate upon the resin wetted web.
U.S. Pat. No. 3,175,331 (Klein) discloses a cleaning and scouring pad comprising one or more fibrous batts, heat-sealed so as to be capable of having enclosed therein a solid washing composition, and in which the outer surface of the pad has grit adhered thereto to provide a continuous, uninterrupted scouring surface extending over the entire outer surface of the pad. A fusible adhesive in liquid form is applied onto either or both surfaces of the fibrous batt sufficient to lightly impregnate the outer surface only of the batt to bond outer fibers together while not filling voids between fibers or penetrating to fibers on the opposite surface of the batt. Therefore, the amount of adhesive is regulated in order to concentrate the adhesive in the area of the surface of the batt instead of the interior of the batt. Where scouring action is desired, abrasive grit is pre-mixed into the impregnating adhesive and applied to the surface(s) of the fibrous batt, such as by spraying. Where the finished pad is to be used for washing instead of scouring, the abrasive material can be omitted from the impregnating resin used in Klein.
However, in order to limit the penetration depth of the liquid resin into the batt as in Klein, careful and time-consuming pre-consideration and monitoring during processing of many parameters, such as resin coating amount, resin flowability, resin viscosity, web thickness, web density, and so forth, would be required.
French Patent Application Publication No. 2,409,095 discloses the use of collapsible, colored foams to concentrate pigments near a surface of a porous support, such as a fibrous support. The pigments or ink base materials are incorporated into and intimately admixed with a binder in dispersion with a mechanical foam machine. A colored foam is formed in which the pigment and binder resins are temporarily suspended within a foam. The resulting colored foam is applied to a fabric surface and subjected to heat to collapse the foam such that the pigment and binder residues only penetrate the fabric surface to a limited degree. French patent 2,409,095 thus allows for surface printing or dyeing of a side or both sides of a fabric without strike-through problems. French patent 2,409,095 teaches pigments, usually relatively light and soft materials, and not dense, solid granulate, as suspended, along with the binder, within and throughout the bulk of the foam. Therefore, pigments located at the lower regions of the foam layer will contact the fabric surface immediately after, or very shortly after, the foam is applied to the fabric surface.
U.S. Pat. No. 4,969,975 (Biggs et al.) describes a process by which a homogeneous sheet comprising a uniform distribution of fibers and/or particles, which otherwise might float and/or settle, by incorporating the fibers and/or particles into a froth or foam itself, and depositing and draining the foamed dispersion on a fibrous support. As the particle additives are distributed within and throughout the foam in Biggs et al., at least a portion of such additives will immediately come into contact with the substrate used as the coating support.
As can be appreciated from the above, there still remains a need for a technique to concentrate abrasive particles in the external surface regions of a foraminous abrasive article, such as a nonwoven, by a simple manufacturing scheme that does not require extensive preparation and monitoring during processing.

Method of making a foraminous abrasive article


A system and method for a sidewall SONOS memory device is provided. An electronic device includes a non-volatile memory. A substrate includes source/drain regions. A gate stack is directly over the substrate and between the source/drain regions. The gate stack has a sidewall. A nitride spacer is formed adjacent to the gate stack. A first oxide material is formed directly adjacent the spacer. An oxide-nitride-oxide structure is formed between the spacer and the gate stack. The oxide-nitride-oxide structure has a generally L-shaped cross-section on at least one side of the gate stack. The oxide-nitride-oxide structure includes a vertical portion and a horizontal portion. The vertical portion is substantially aligned with the sidewall and located between the first oxide material and the gate sidewall. The horizontal portion is substantially aligned with the substrate and located between the first oxide and the substrate.

Non-volatile memory has been integrated into a wide range of electronic technologies, including cellular communication, digital video, digital audio, and compact data storage (i.e., flash memory cards, flash memory sticks, and USB flash drives). Silicon-oxide-nitride-oxide-silicon (SONOS) memory is a non-volatile semiconductor memory, at the heart of which is a transistor that can retain a value without a constant power supply. During operation, a conventional SONOS memory transistor stores a single charge in a nitride layer sandwiched between two thin oxide layers. The oxide layers are in turn sandwiched between two silicon layers. These two silicon layers are typically a polysilicon gate electrode (i.e., the controlling gate) and the silicon substrate.
Sidewall SONOS technology improves upon SONOS technology by storing two bits per transistor. A sidewall SONOS transistor stores two discrete bits, in contrast with the one bit stored by a conventional SONOS transistor. FIG. 1 shows the cross-section of a known sidewall SONOS transistor 10. The transistor 10 includes an oxide 16 formed around a gate electrode 12 and a gate dielectric 14. The gate dielectric 14 is directly over a silicon substrate 17. A material 18 with charge trapping properties overlies the substrate 17, and includes portions 22 that are directly adjacent the oxide 16. A silicon dioxide spacer 20 is adjacent the gate electrode 12.
The sidewall SONOS transistor 10 is repeated thousands, millions, and billions of times in a memory array (not shown). Because the sidewall SONOS transistor 10 is repeated numerous times, the design of the sidewall SONOS transistor 10 is critical to the reliability and compactness of the SONOS memory as a whole.
The sidewall SONOS transistor 10 in FIG. 1 has several disadvantages. First, the sidewall SONOS transistor 10 may suffer from electron drift. During operation, an electric charge is stored in the region 22. However, the region 22 is electrically connected to larger adjacent regions of the charge storing material 18, and electrons (not shown) in the region 22 may drift, thus degrading the charge density in the region 22. Charge density in the region 22 is critical to the performance of the sidewall SONOS transistor 10. The degradation of electric charge density in the region 22 may erroneously alter the value read from the transistor 10. The flaw is compounded by the number of times the sidewall SONOS transistor 10 is repeated in the SONOS memory.
In addition to suffering from electron drift, the sidewall SONOS transistor 10 shown in FIG. 1 is poorly suited for downward scalability. For example, the transistor 10 may not be scalable to a semiconductor process technology generation below 0.130 microns (μm). The transistor 10 has a silicon dioxide (SiO2) spacer 20, but technological barriers exist, which prevent the manufacturing of SiO2 gate spacers in sub-0.130 μm semiconductor manufacturing technologies. In addition, the substantially L-shaped formation of the region 22 of the charge trapping material 18 is not reproducible in a sub-0.130 μm manufacturing technology.
FIG. 2 shows a cross-section of a conventional sidewall SONOS transistor 30 manufactured in a sub-0.130 μm technology generation. The transistor 30 includes a gate spacer 32, separated from the gate electrode 12 and gate dielectric 14, by an oxide 16. The spacer 32 has charge storing properties, and during operation an electric charge may be formed in the spacer 32. However, the spacer 32 is sufficiently voluminous to suffer from electron drift. The dissipation of electrons in the spacer 32 reduces the charge density in the spacer 32, resulting in possible data retention and reliability issues.
Hence, there is a need for a reliable SONOS transistor that is scalable to sub-0.130 μm semiconductor manufacturing technology generations.

Structure and method for a sidewall SONOS memory device

Taiwan Semiconductor Manufacturing Company, Ltd.


A nucleic acid includes the 5′ untranslated region, a virus protein-coding region including the NS3 protein coding sequence, the NS4A protein coding sequence, the NS4B protein coding sequence, the NS5A protein coding sequence, and the NS5B protein coding sequence, and the 3′ untranslated region of the HCV J6CF genome in that order from the 5′ to 3′ direction. The NS4A protein coding sequence has a mutation for substituting alanine at position 1680 with glutamic acid, as determined on the basis of the amino acid sequence of the precursor protein of the J6CF strain.

The hepatitis C virus (hereinafter, referred to as HCV) is a virus belonging to the flavivirus family the genome of which is a single-stranded (+) sense RNA and is known to cause hepatitis C. Based on recent studies, it has been revealed that HCV is classified into many types depending on genotype or serotype. According to phylogenetic analysis by Simmonds et al. using nucleotide sequences of HCV strains, HCV genotypes are classified into six types, and each type is further classified into several subtypes (Simmonds et al., Hepatology, (1994) Vol. 10, pp. 1321-1324). The full-length genomic nucleotide sequences of a plurality of HCV genotypes have also been determined to date (Choo et al., Science, (1989) Vol. 244, pp. 359-362, Kato et al., J. Med. Virol., (1992) Vol. 64, pp. 334-339, Okamoto et al., J. Gen. Virol., (1992) Vol. 73, pp. 673-679, Yoshioka et al., Hepatology, (1992) Vol. 16, pp. 293-299 and Mori et al., Biochem. Biophys. Res. Commun., (1992) Vol. 183, pp. 334-342).
Until recently, infection of cultured cells with HCV and replication of HCV genomes in cultured cells have been impossible. Accordingly, studies on mechanisms of HCV replication and infection have required in vivo experiments using chimpanzees as experimental animals. However, subgenomic replicon RNAs have been produced from the Con1 strain, the HCV-N strain, and the HCV-O strain belonging to the HCV genotype 1b, and the H77c strain belonging to the HCV genotype 1a, and this enabled studies on the HCV replication mechanism via in vitro experiments using cultured cells (JP 2001-17187 A, Lohmann et al., Science, (1999) Vol. 285, pp. 110-113, Blight et al., Science, (2000) Vol. 290, pp. 1972-1974, Friebe et al., J. Virol., (2001) Vol. 75, pp. 12047-12057 and Ikeda et al., J. Virol., (2002) Vol. 76, pp. 2997-3006). Herein, the subgenomic replicon RNA of HCV means an RNA which comprises a portion of HCV genome necessary for replication, and can autonomously replicate an RNA derived from the HCV genome when introduced into cultured cells, but does not have an ability to produce infectious HCV particles.
In addition, subgenomic replicon RNAs, and full-genomic replicon RNAs producing infectious HCV particles when introduced into Huh7 cells in vitro have been produced from the JFH-1 strain belonging to the HCV genotype 2a, and this enabled studies on the HCV infection mechanism via in vitro experiments using cultured cells (Kato et al., Gastroenterology, (2003) Vol. 125, pp. 1808-1817 and Wakita et al., Nature Medicine, (2005) Vol. 11, pp. 791-796). Herein, the full-genomic replicon RNA of HCV means an RNA which comprises a full-length HCV genome, i.e., a 5′ untranslated region, structural genes, non-structural genes, and a 3′ untranslated region, and can autonomously replicate an RNA derived from the HCV genome when introduced into cultured cells.
The J6CF strain is a HCV of genotype 2a, as is the JFH-1 strain. Homology between the J6CF strain and the JFH-1 strain is as high as about 90% at nucleic acid level and 91% at amino acid level. However, the J6CF strain lacks replication ability of its subgenomic replicon RNA, and it is not capable of virus particle production in a cell culture system using Huh7 cells.
Recently, substitution of an NS3 protein coding region, an NS5B protein coding region, and a 3′ untranslated region of the J6CF genome with the sequences of the same regions of the JFH-1 strain was found to lead to autonomous replication in Huh7 cells (Murayama et al., J. Virol., (2007) Vol. 81, pp. 8030-8040). In addition, the subgenomic replicon RNA and full-genomic replicon RNA prepared by substituting an NS3 protein coding region and a 3′ untranslated region of the J6CF genome with the sequences of the same regions of the JFH-1 strain and introducing three adaptive mutations in the NS5B protein coding region have been demonstrated to have autonomous replication ability and virus particle-producing ability (Murayama et al., PLoS Pathogens., (2010) Vol. 6, e1000885).
An experimental system that can efficiently amplify viruses is indispensable for studies on HCV and research and development of anti-HCV drugs. A system for amplifying HCV in cultured cells and a system for evaluating the propagation of HCV in cultured cells enable efficient screening of anti-HCV drugs. In addition, based on the systems, mechanisms of virus propagation will be understood, and new targets of anti-HCV drugs will be also found, and development of innovative anti-HCV drugs against such targets can be expected. Production of replicon RNA capable of autonomous replication in cultured cells or virus particle production is also desired for the J6CF strain.
Up to the present, however, it is believed that a full-genomic replicon RNA having infectious HCV particle-producing ability is required to comprise a sequence derived from the JFH-1 strain as a non-structural gene that is indispensable for virus replication. J6CF strain-derived full-genomic replicon RNAs autonomously replicating in cultured cells and producing HCV particles have been obtained, but they are limited to those composed of chimeric nucleic acids resulting from substitution of a considerable part of the non-structural genes in the J6CF genome with a sequence of the same region derived from the JFH-1 strain. Full-genomic replicon RNA having the J6CF genome as a backbone and having infectious HCV particle-producing ability is necessary for research of the J6CF strain, but has not yet been obtained.
As described above, subgenomic replicon RNA or full-genomic replicon RNA derived from HCV of genotype 2a corresponding to the replication mechanisms or replication efficiency of HCV strains other than the JFH-1 strain has not yet been obtained. Accordingly, it has been difficult to identify factors necessary for HCV replication serving as targets of new anti-HCV drugs or to screen for anti-HCV drugs capable of exerting drug efficacy independent of the replication mechanisms or replication efficiency. In addition, the types of artificially producible HCV particles serving as a raw material for HCV vaccine have been limited.
It could therefore be helpful to provide a replicon RNA having the genome of the hepatitis C virus J6CF strain as a backbone and having autonomous replication ability.

Mutant replicon derived from genome of hepatitis C virus J6CF strain

Japan As Represented by Director-General of National Institute of Infectious Diseases


A CMOS image sensor (CIS) device includes an array of pixels, each pixel including a sensing element (e.g., a photodiode) and access circuitry. To facilitate the passage of light to the photodiode, each pixel includes a via wave guide (VWG) defined in the metallization layer formed over the pixel's photodiode. The VWG includes an upper light concentrator having a curved (e.g., parabolic) surface extending from a relatively wide upper opening to a relatively small lower opening. The VWG also includes a lower section extending between the lower opening of the light concentrator and the associated photodiode. A mirror coating is optionally formed on the surface of the VWG. An optional light-guiding material and/or color filter materials are disposed inside the VWG. An optional microlens is formed over the VWG.

Solid state image sensors are used, for example, in video cameras, and are presently realized in a number of forms including charge-coupled devices (CCDs) and CMOS image sensors (CISs). CISs sensors are based on a two dimensional array of pixels that are fabricated using CMOS fabrication techniques. Each CIS pixel includes a sensing element (e.g., a photodiode) and access circuitry that are fabricated on a semiconductor substrate, and connected to control circuits by way of metal address and signal lines. These metal lines are supported in insulation material that is deposited over the upper surface of the semiconductor substrate, and positioned along the peripheral edges of the pixels to allow light to pass between the metal lines to the sensing elements through the insulation material. In color image sensors, each pixel also includes a color filter located over the sensing element. An array of microlenses is sometimes located over the metallization layer to focuses light from an optical image through the color filter and the insulation material into the image sensing elements. Each image sensing element is capable of converting a portion of the optical image passed by the color filter into an electronic signal. The electronic signals from all of the image sensing elements are then used to regenerate the optical image on, for example, a video monitor.
The quality of an image generated by a conventional CIS is at least in part determined by the amount of light that reaches the photodiode of each pixel. As indicated above, the photodiode of each pixel covers only a portion of the entire pixel area, with the access circuitry and address/signal lines taking up the remaining CIS surface area. Accordingly, in the absence of microlenses, only a portion of the light incident on the upper surface of the CIS is captured by the photodiodes. Further, when color filters are present, only a portion of the light directed toward a particular photodiode is passed by the color filter, further reducing the amount of captured light that can be used to generate image information. Moreover, because the light must pass through the semi-opaque insulation material of the metallization layer, a portion of the filtered light directed toward each photodiode is reflected or refracted away from the photodiode. Some of this reflected/refracted light may strike an adjacent photodiode, producing blurring and/or inaccurate image color.
What is needed is a CIS that facilitates enhanced image detection by providing a structure for capturing and concentrating substantially all of the light incident on the CIS, and directing the concentrated light onto the CIS's photodiodes.

Via wave guide with curved light concentrator for image sensing devices


‘LIRF’ is a distinctive variety of Liriope muscari which is characterized by the combination of its narrow leaf width, dense foliage and a pink flower color.

The present invention relates to a new and distinct perennial variety of Liriope muscari, which has been given the variety denomination of 'LIRF'. Its market class is that of an ornamental plant. 'LIRF' is intended for use in landscaping and as a decorative plant.
The Liriope muscari variety 'LIRF' was finally selected in 2005 in an Australian nursery in the state of New South Wales following a selection process carried out from 2002 to 2005 involving approximately 20000 seedlings of Liriope muscari cultivar 'Big Blue' (unpatented) production stock. 'LIRF' is a seedling selection from open pollinated Liriope muscari cultivar 'Big Blue'. 'LIRF' was selected due to its narrow leaf width combined with dense foliage and a pink flower color. 'LIRF' was first propagated asexually by division in the state of New South Wales, Australia and has since been asexually propagated by division and micropropagation. The distinctive characteristics of the inventive 'LIRF' variety are stable from generation to generation; clones of the variety produced by asexual reproduction maintain the distinguishing characteristics of the original plant.
'LIRF' has an upright growth habit with a very dense shoot density whereas 'Big Blue' has a semi upright growth habit and a medium shoot density. 'LIRF' has a narrow leaf width resulting in finer looking foliage whereas 'Big Blue' has a broad leaf width. 'LIRF' has a pink flower color whereas 'Big Blue' has a light purple to lilac flower color.
An application for plant breeders' rights for variety 'LIRF' has been lodged with the Australian Plant Breeders' Rights Office, and was first gazetted in Mar. 27, 2006 under Application No. 2006/038.


An original scanning apparatus comprising original support means on which an original is rested, elongated light source means for illuminating the original rested on the original support means, reflection means for reflecting light from the elongated light source means toward the original support means. The elongated light source means is fixed and the reflection means being shiftable, so that the original rested on the original support means is scanned by shifting the reflection means. Apparatus further includes side reflector means disposed at a longitudinal end of the elongated light source means and extending along a shifting direction of the reflection means.

1. Field of the Invention
The present invention relates to an original scanning apparatus used with a copying machine, image reader and the like, and more particularly, it relates to an apparatus for scanning an original by shifting a reflection member with respect to a fixed original support plate and a fixed light source.
2. Related Background Art
Conventional slit exposure apparatus wherein an original support plate and a light source are fixed are already known, as disclosed in the Japanese Patent Laid-open Nos. 54-133140, 54-42129 and the like. FIGS. 18 and 19 show examples of conventional illuminating or lighting equipment.
The lighting equipment shown in FIG. 18 includes an elongated light source 101 disposed at one side of an original. The lighting equipment further includes reflection mirrors 102a, 102b for reflecting light from the elongated light source. The reference numeral 103 denotes an original support plate; 104 denotes an original; and 105 denotes a shiftable reflection member for optically scanning the original 104 by shifting the light from the reflection mirror 102b horizontally. On the other hand, the lighting equipment shown in FIG. 19 includes light sources disposed at both sides of an original. The same constructural elements as those shown in FIG. 18 are designated by the same reference numerals.
By the way, in the above-mentioned conventional lighting equipments, there arose a problem that the brightness on a peripheral portion of the original, particularly on a lateral edge portion of the original extending along the elongated light source was decreased as the shiftable reflection member was shifted away from the elongated light source because of the increased distance between the light source and a slit exposure scanning position.
FIG. 20 shows the distribution of the brightness on the original when the original is illuminated from one side thereof as in the equipment of FIG. 18, and FIG. 21 shows the distribution of the brightness on the original when the original is illuminated from both sides thereof as in the equipment of FIG. 19. In both cases, the distribution of the brightness is shows as the equi-illumination curves or lines. In FIGS. 20 and 21, arrows a and b show a condition that the light beams emitted laterally from the light source 101 escape outwardly from the surface of the original, and the thick arrow indicates a low illumination portion.
The reduction in the intensity of illumination as mentioned above causes the fog phenomenon in the outputted image from the copying machine and the like, thus worsening the ability to copy with high quality. Further, since the light source is fixed, the light source can more easily be arranged near the original support plate in comparison with the case where the light source is shiftable. However, when the light source is an elongated light source comprised of a plurality of illuminous bodies arranged in a line, the unevenness in the amount of light (intensity of illumination) is generated on the surface of the original in correspondence with the pitch between the adjacent luminous bodies, and, thus, it is impossible to uniformly illuminate the original along the longitudinal direction of the elongated light source.

Original scanning apparatus with fixed light source


A system and method that automatically resolves conflicts among sensor information in a sensor fusion robot system. Such methods can accommodate converging ambiguous and divergent sensor information in a manner that can allow continued, and relatively accurate, robotic operations. The processes can include handling sensor conflict via sensor prioritization, including, but not limited, prioritization based on the particular stage or segment of the assembly operation when the conflict occurs, overriding sensor data that exceeds a threshold value, and/or prioritization based on evaluations of recent sensor performance, predictions, system configuration, and/or historical information. The processes can include responding to sensor conflicts through comparisons of the accuracy of workpiece location predictions from different sensors during different assembly stages in connection with arriving at a determination of which sensor(s) is providing accurate and reliable predictions.

A variety of operations can be performed during the final trim and assembly (FTA) stage of automotive assembly, including, for example, door assembly, cockpit assembly, and seat assembly, among other types of assemblies. Yet, for a variety of reasons, only a relatively small number of FTA tasks are typically automated. For example, often during the FTA stage, while an operator is performing an FTA operation, the vehicle(s) undergoing FTA is/are being transported on a line(s) that is/are moving the vehicle(s) in a relatively continuous manner. Yet such continuous motions of the vehicle(s) can cause or create certain irregularities with respect to at least the movement and/or position of the vehicle(s), and/or the portions of the vehicle(s) that are involved in the FTA. Moreover, such stop and go motion can cause the vehicle to be subjected to movement irregularities, vibrations, and balancing issues during FTA, which can prevent, or be adverse to, the ability to accurately model or predict the location of a particular part, portion, or area of the vehicle that directly involved in the FTA. Further, such movement irregularities can prevent the FTA from having a consistent degree of repeatability in terms of the movement and/or positioning of each vehicle, or its associated component, as each subsequent vehicle and/or component passes along the same area of the assembly line. Accordingly, such variances and concerns regarding repeatability can often preclude the use of traditional teach and repeat position based robot motion control in FTA operations.
Accordingly, although various robot control systems are available currently in the marketplace, further improvements are possible to provide a system and means to calibrate and tune the robot control system to accommodate such movement irregularities.

Automatic sensor conflict resolution for sensor fusion system


The invention relates to protein complexes and fusion proteins including at least two different cytokine molecules. The protein complexes and fusion proteins may further include a targeting moiety such as a region of an immunoglobulin. Methods of using the protein complexes and fusion proteins are also disclosed.

The regulatory networks controlling the immune system rely on secreted protein signaling molecules termed cytokines to turn on and off the functions of immune cells as well regulate their proliferation. These responses generally involve multiple cytokines that act in concert to achieve the desired biological effect. Certain cytokines such as interleukin-2 (IL-2) can induce immune cell proliferation by themselves and can activate other functions including secondary cytokine secretion. Another cytokine, interleukin-12 (IL-12) [reviewed by Trinchieri, 1994, Blood 84:4008–4027], can induce proliferation of certain immune cells and induce another key immune modulator, interferon-γ (IFN-γ). This induction of IFN-γ is a key activity of IL-12, although IL-12 has other important activities that are IFN-γ independent. Since IL-12 itself is induced at an early stage in infectious disease situations, it is thought to link the innate and acquired immune systems.
Many in vitro studies with both mouse and human immune cells have shown the importance of cytokine combinations in the development of optimal immune responses. For example, most T cells do not express IL-12 receptors (IL-12R) until they have been activated with mitogens or cultured in high concentrations of IL-2 [Desai et al. (1992), J. Immunol. 148:3125–3132]. Once the receptors are expressed, the cells become far more responsive to IL-12. Furthermore, IL-12 induces IFN-γ transcription, but IFN-γ mRNA is degraded shortly thereafter, in the presence of IL-2, the mRNA is stabilized, resulting in a dramatic increase in the amount of IFN-γ produced [Chan et al. (1992) J. Immunol. 0.148:92–98]. In other studies, it was found that the cytokine combinations IL-3 plus IL-11 or IL-3 pus Steel Factor had a synergistic effect with IL-12 on the proliferation of early hematopoietic progenitor cells [Trinchieri, 1994; cited above]. The combination of interleukin-4 and GM-CSF is particularly useful in stimulating dendritic cells (Palucka et al. [1998] J. Immunology 160:4587–4595). For stimulation of the cell-mediated immune response, it is also useful to combine IL-12 with IL-18, a recently discovered Th1-promoting cytokine with some activities that are complementary to IL-12 (Hashimoto et al. [1999] J. Immunology 163:583–589; Barbulescu et al. [1998] J. Immunology 160:3642–3647). In addition, IL-2 and interferon-γ are synergistic in certain circumstances [Palladino, M. A., U.S. Pat. No. 5,082,658].
In many of these synergy studies it was found that the relative level of each cytokine was very important. Whereas the addition of IL-12 in the presence of suboptimal amounts of IL-2 led to synergy in the induction of proliferation, cytolytic activity and IFN-γ induction, combinations of IL-2 and IL-12 using a high dose of one cytokine were found to be antagonistic [Perussia et al., J. Immunol. 149:3495–3502 (1992); Mehrotra et al., J. Immunol. 151:2444–2452 (1993)]. A similar situation also exists in combinations of IL-12 and IL-7.
Synergy studies between IL-12 and other cytokines for the generation of anti-tumor responses in mice have also shown mixed results. In some models synergy was seen at suboptimal doses of each cytokine and higher doses led to enhanced toxicity, while in other models, combinations of IL-12 and IL-2 showed little or no synergy [see, for example, Nastala et al., J. Immunol. 153:1697–1706. (1994)]. These results may reflect the inherent difficulty of combining two potentially synergistic agents in vivo, especially when there is the need to maintain a fixed ratio of activities of two agents with different pharmacological properties, such as different circulating half-life and biodistribution.
In in vitro cell culture experiments, it is straightforward to control cytokine levels, but many factors can affect the relative biodistribution and localization of cytokines in vivo, thus affecting their immunostimulatory capacity. The most important of these factors is the half-life. The half-life of IL-2 in the circulation after bolus injection is approximately 10 minutes. In striking contrast to these pharmacokinetic properties, the circulating half-life of IL-12 has been reported to be >3 hr in mice [Wysocka et al (1995) Eur. J. Immunol. 25:672] and from 5–10 hr in humans [Lotze et al. (1996) Ann NY Acad Sci 795:440–454].
This difference is thought to be due to the relatively small sizes of both IL-2 and GM-CSF (15–25 kD vs. 75 kD for IL-12), allowing IL-2 and GM-CSF to be cleared by renal filtration. Proteins with a molecular weight of less than about 50 kD are cleared by renal filtration. Almost all cytokines are smaller than 50 kD and undergo similar, rapid clearance by renal filtration. When treatment with two such small, rapidly cleared cytokines is desired, it is sufficient to simply co-administer the cytokines. However, co-administration is not optimal for cytokines with significantly different half-lives.
The systemic administration of cytokines is difficult due to their deleterious side effects. For example, high levels of Interferon-alpha result in significant side effects, including skin, neurologic, immune and endocrine toxicities. It is expected that multiple cytokine fusions might show particularly serious side effects.
To reduce side effects of systemic administration of cytokines, one strategy is to fuse a cytokine to a second molecule with targeting capability. Fusions in which an Fc region is placed at the N-terminus of a another protein (termed 'immunofusins' or 'Fc-X' fusions, where X is a ligand such as Interferon-alpha) have a number of distinctive, advantageous biological properties [Lo et al., U.S. Pat. Nos. 5,726,044 and 5,541,087; Lo et al., Protein Engineering 11:495]. In particular, such fusion proteins can still bind to the relevant Fc receptors on cell surfaces. However, when the ligand binds to its receptor on a cell surface, the orientation of the Fc region is altered and the sequences that mediate antibody-dependent cell-mediated cytotoxicity (ADCC) and complement fixation appear to be occluded. As a result, the Fc region in an Fc-X molecule does not mediate ADCC or complement fixation effectively. The cytotoxic effect due to the fusion of an N-terminal cytokine and a C-terminal Fc region is well known. For example, fusion of IL-2 to the N-terminus of an Fc region creates a molecule that is able to bind to cells bearing the IL-2 receptor, fix complement, and lyse the cells as a result [Landolfi, N. F. (1993) U.S. Pat. No. 5,349,053]. In contrast, Fc-IL-2 fusion proteins do not have this property. Thus, Fc-X fusions are expected to have the virtues of increased serum half-life and relative concentration in the liver, without the deleterious effects of ADCC and complement fixation.
It has been demonstrated that many different proteins with short serum half-lives can be fused to an Fc region in an Fc-X configuration, and the resulting fusions have much longer serum half-lives. However, the serum half-lives of two different Fc fusions will not generally be identical. Thus, when delivery of two different X moieties is desired, co-administration of two different Fc-X proteins will not generally be optimal.
Under some circumstances, a better approach is to target the effect of the cytokine to a cell surface antigen by fusing it to an antibody (or fragment derived therefrom) having specificity and affinity for that antigen (Gillies, U.S. Pat. No. 5,650,150; Gillies et al., Proc. Natl. Acad. Sci. 89:1428) or by linking a protein antigen and stimulatory cytokine via a peptide linkage in the form of a fusion protein (Hazama et al, Vaccine 11:629). While antibodies themselves can increase the half-life of a fused cytokine, there are still differences between different cytokine fusions with the same antibody [see, for example, Gillies et al., Bioconjugate Chem. 4:230–235 (1993); Gillies et al., J. Immunol. 160:6195–6203] that would make co-localization at a target site difficult. As discussed above, this could lead to an imbalance in cytokine activities and decrease the desired synergistic effects. In addition, the use of two different fusion proteins requires testing each fusion separately for its safety and effectiveness profile, and then further testing as mixtures.

This patent is subject to a terminal disclaimer.