Abstract: Provided herein are alphavirus vectors derived from South African Arbovirus No. 86 (S.A.AR86) comprising attenuating mutations and methods of making the same. Also provided are improved viral vectors and helper constructs comprising a S.A.AR86 capsid enhancer sequence. The present invention also provides S.A.AR86 replicon and helper constructs comprising an alphavirus capsid enhancer sequence. Further provided are methods of administering an alphavirus vector comprising a heterologous nucleotide sequence (preferably encoding an immunogen or a therapeutic polypeptide) according to the invention to a cell or subject. In preferred embodiments, the alphavirus vector delivers the heterologous nucleotide sequence to the cells of the bone, bone marrow, and/or bone-associated connective tissue.

Abstract: The present invention provides a helper cell for expressing an infectious, replication defective, alphavirus particle in an alphavirus-permissive cell. The helper cell includes (a) a first helper RNA encoding (i) at least one alphavirus structural protein, and (ii) not encoding at least one alphavirus structural protein; and (b) a second helper RNA separate from the first helper RNA, the second helper RNA (i) not encoding the alphavirus structural protein encoded by the first helper RNA, and (ii) encoding the at least alphavirus one structural protein not encoded by the first helper RNA, such that all of the alphavirus structural proteins assemble together into alphavirus particles in the cell. Preferably, the helper cell also includes a replicon RNA encoding an alphavirus packaging sequence and an inserted heterogeneous RNA.

Abstract: A diversity receiver circuit system (10) including a primary channel (20) and a diversity channel (22), where analog input signals are converted to differential signals in both channels (20, 22). The receiver circuit system (10) includes a multiplexer (14) and a variable gain amplifier (12) formed on a single RF integrated circuit chip (16), where the multiplexer (14) is positioned before the amplifier (12). The differential signals in the primary channel (20) and the diversity channel (22) are applied to an amplified path (72, 78) and a non-amplified path (76, 82) in the multiplexer (14). A control signal selects one of the amplified primary channel signal, the non-amplified primary channel signal, the amplified diversity channel signal or the non-amplified diversity channel signal.

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

Abstract: The present invention provides a helper cell for expressing an infectious, replication defective, alphavirus particle in an alphavirus-permissive cell. The helper cell includes (a) a first helper RNA encoding (i) at least one alphavirus structural protein, and (ii) not encoding at least one alphavirus structural protein; and (b) a second helper RNA separate from the first helper RNA, the second helper RNA (i) not encoding the alphavirus structural protein encoded by the first helper RNA, and (ii) encoding the at least one alphavirus structural protein not encoded by the first helper RNA. Preferably, the helper cell is co-transfected with a replicon RNA encoding an alphavirus packaging segment and an inserted heterogeneous RNA, such that all of the alphavirus structural proteins assemble together into alphavirus particles in the cell, with said replicon RNA packaged therein.

Abstract: The present invention provides a helper cell for expressing an infectious, replication defective, alphavirus particle in an alphavirus-permissive cell. The helper cell includes (a) a first helper RNA encoding (i) at least one alphavirus structural protein, and (ii) not encoding at least one alphavirus structural protein; and (b) a second helper RNA separate from the first helper RNA, the second helper RNA (i) not encoding the alphavirus structural protein encoded by the first helper RNA, and (ii) encoding the at least alphavirus one structural protein not encoded by the first helper RNA, such that all of the alphavirus structural proteins assemble together into alphavirus particles in the cell. Preferably, the helper cell also includes a replicon RNA encoding an alphavirus packaging sequence and an inserted heterogeneous RNA.

Abstract: The present invention provides a helper cell for expressing an infectious, replication defective, alphavirus particle in an alphavirus-permissive cell. The helper cell includes (a) a first helper RNA encoding (i) at least one alphavirus structural protein, and (ii) not encoding at least one alphavirus structural protein; and (b) a second helper RNA separate from the first helper RNA, the second helper RNA (i) not encoding the alphavirus structural protein encoded by the first helper RNA, and (ii) encoding the at least one alphavirus structural protein not encoded by the first helper RNA. Preferably, the helper cell is co-transfected with a replicon RNA encoding an alphavirus packaging segment and an inserted heterogeneous RNA, such that all of the alphavirus structural proteins assemble together into alphavirus particles in the cell, with said replicon RNA packaged therein.

Abstract: The present invention provides a helper cell for expressing an infectious, replication defective, alphavirus particle in an alphavirus-permissive cell. The helper cell includes (a) a first helper RNA encoding (i) at least one alphavirus structural protein, and (ii) not encoding at least one alphavirus structural protein; and (b) a second helper RNA separate from the first helper RNA, the second helper RNA (i) not encoding the alphavirus structural protein encoded by the first helper RNA, and (ii) encoding the at least one alphavirus structural protein not encoded by the first helper RNA. Preferably, the helper cell is co-transfected with a replicon RNA encoding an alphavirus packaging segment and an inserted heterogeneous RNA, such that all of the alphavirus structural proteins assemble together into alphavirus particles in the cell, with said replicon RNA packaged therein.

Abstract: The present invention provides a helper cell for expressing an infectious, replication defective, alphavirus particle in an alphavirus-permissive cell. The helper cell includes (a) a first helper RNA encoding (i) at least one alphavirus structural protein, and (ii) not encoding at least one alphavirus structural protein; and (b) a second helper RNA separate from the first helper RNA, the second helper RNA (i) not encoding the alphavirus structural protein encoded by the first helper RNA, and (ii) encoding the at least alphavirus one structural protein not encoded by the first helper RNA, such that all of the alphavirus structural proteins assemble together into alphavirus particles in the cell. Preferably, the helper cell also includes a replicon RNA encoding an alphavirus packaging sequence and an inserted heterogeneous RNA.

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

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

Abstract: The present invention provides a helper cell for expressing an infectious, replication defective, alphavirus particle in an alphavirus-permissive cell. The helper cell includes (a) a first helper RNA encoding (i) at least one alphavirus structural protein, and (ii) not encoding at least one alphavirus structural protein; and (b) a second helper RNA separate from the first helper RNA, the second helper RNA (i) not encoding the alphavirus structural protein encoded by the first helper RNA, and (ii) encoding the at least alphavirus one structural protein not encoded by the first helper RNA, such that all of the alphavirus structural proteins assemble together into alphavirus particles in the cell. Preferably, the helper cell also includes a replicon RNA encoding an alphavirus packaging sequence and an inserted heterogeneous RNA.

Abstract: A method of protecting a subject against a disease comprises administering a recombinant Venezuelan Equine Encephalitis (VEE) virus to the subject in an effective immunogenic amount, with the VEE virus containing a heterologous DNA segment, and with the heterologous DNA segment comprising a promoter operable in the subject operatively associated with a DNA encoding an immunogenic protein or peptide effective for protecting the subject from the disease. Preferred promoters are VEE 26S subgenomic promoters, and preferred immunogens are viral immunogens. Novel attenuating mutations useful in carrying out the invention are also disclosed.

Abstract: The present invention provides a recombinant DNA comprising a cDNA coding for an infectious South African Arbovirus No. 86 (S.A.AR86) virus RNA transcript and a heterologous promoter positioned upstream from the cDNA and operatively associated therewith. The present invention also provides an infectious RNA transcript encoded by the cDNA, and infectious attenuated viral particles containing the RNA transcript encoded by the cDNA.

Abstract: Novel attenuating mutations of Venezuelan Equine Encephalitis (VEE) are disclosed. Further aspects of the invention include an infectious VEE virus transcript encoded by cDNA clones, infectious VEE virus particles, and pharmaceutical formulations containing such infectious particles. Also disclosed are recombinant VEE virus containing a heterologous RNA segment.

Abstract: A fiberboard pallet comprises a top deck of multi-wall corrugated fiberboard having a lower surface to which three, four or more elongated legs are permanently attached, for example, by gluing. Each leg is formed of a die cut blank having a series of five panels which are folded into a quadrilateral shape to form an initially open elongated passage. End flaps at the ends of each leg are folded and overlap each other to close the ends. At least two side openings are formed in each leg and any communication into the passage of the quadrilateral shape is closed by closing flaps formed as one piece with side panels of each leg. A bottom deck is fixed to the bottom of the legs.

Abstract: A fiberboard pallet comprises a deck of multi-wall corrugated fiberboard having a lower surface to which three, four or more elongated legs are permanently attached, for example, by gluing. Each leg is formed of a die cut blank having a series of five panels which are folded into a quadrilateral shape to form an initially open elongated passage. End flaps at the ends of each leg are folded and overlap each other to close the ends. At least two side openings are formed in each leg and any communication into the passage of the quadrilateral shape is closed by closing flaps formed as one piece with side panels of each leg.

Abstract: Apparatus and method to move granular materials in an even flow by centrifugal force from a rotating surface to a plurality of circumferentially arranged receiving points includes a rotatable inverted cone provided with a peripheral base flange and having an angle of inclination of the cone surface matching the angle of repose for the material to be distributed. Material delivered to the apex of the cone flows by gravity down the cone surface to establish the angle of repose for such material with a toe point at the flange where it comes to rest when the cone is stationary. A cylindrical hopper, to accumulate material from the cone, is vertically adjustably secured to the flange so that the bottom of the hopper is spaced above the flange to define a flow passageway which can be enlarged or reduced by vertical adjustment of the hopper relative to the flange.

Abstract: A fiberboard pallet comprises a deck of multi-wall corrugated fiberboard having a lower surface to which three, four or more elongated legs are permanently attached, for example, by gluing. Each leg is formed of a die cut blank having a series of five panels which are folded into a quadrilateral shape to form an initially open elongated passage. End flaps at the ends of each leg are folded and overlap each other to close the ends. At least two side openings are formed in each leg and any communication into the passage of the quadrilateral shape is closed by closing flaps formed as one piece with side panels of each leg.

Abstract: A DNA comprises a cDNA clone coding for an infectious Venezuelan Equine Encephalitis Virus RNA transcript and a heterologous promoter sequence positioned upstream from the cDNA clone and operatively associated therewith. A method of making a live attenuated Togavirus useful as a vaccine, and cDNA clones which code for attenuated Togaviruses, is also disclosed.