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Powell and Richard H. To isolate the laser system from vibration, the foundation of each laser bay was made independent of the rest of the structure. If the compositions of the invention are to be administered topically, the compositions can be formulated in the form of an ointment, cream, transdermal patch, lotion, gel, shampoo, spray, aerosol, solution, emulsion, or other form well known to one of skill in the art. Wikimedia Commons has media related to National Ignition Facility. Girl, Wash Your Face:

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Grateful American: Addition of hyaluronidase in the injectable solution improves human bioavailability following parenteral administration, particularly subcutaneous administration. It also allows for greater injection site volumes i. The compositions of this invention may be in a variety of forms. These include, for example, liquid, semi-solid and solid dosage forms, such as liquid solutions e.

The particular form depends on the intended mode of administration and therapeutic application. Typical particular compositions are in the form of injectable or infusible solutions, such as compositions similar to those used for passive immunization of humans with other antibodies.

A particular mode of administration is parenteral e. In a particular embodiment, the antibody is administered by intravenous infusion or injection. In another particular embodiment, the antibody is administered by intramuscular or subcutaneous injection. Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage.

The composition can be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable to high drug concentration. Sterile injectable solutions can be prepared by incorporating the active compound i.

Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile, lyophilized powders for the preparation of sterile injectable solutions, particular methods of preparation are vacuum drying and spray-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

The proper fluidity of a solution can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prolonged absorption of injectable compositions can be brought about by including, in the composition, an agent that delays absorption, for example, monostearate salts and gelatin.

In certain embodiments, the active compound may be prepared with a carrier that will protect the compound against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems.

Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art. Robinson, ed. In certain embodiments, an antibody or antibody portion of the invention may be orally administered, for example, with an inert diluent or an assimilable edible carrier.

For oral therapeutic administration, the compounds may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.

To administer a compound of the invention by other than parenteral administration, it may be necessary to coat the compound with, or co-administer the compound with, a material to prevent its inactivation. Supplementary active compounds can also be incorporated into the compositions.

Furthermore, one or more antibodies of the invention may be used in combination with two or more of the foregoing therapeutic agents. Such combination therapies may advantageously utilize lower dosages of the administered therapeutic agents, thus avoiding possible toxicities or complications associated with the various monotherapies.

In certain embodiments, an antibody to RAGE or fragment thereof is linked to a half-life extending vehicle known in the art.

Such vehicles include, but are not limited to, the Fc domain, polyethylene glycol, and dextran. Such vehicles are described, e.

Application Serial No. In a specific embodiment, nucleic acid sequences comprising nucleotide sequences encoding an antibody of the invention or another prophylactic or therapeutic agent of the invention are administered to treat, prevent, manage, or ameliorate a disorder or one or more symptoms thereof by way of gene therapy.

Gene therapy refers to therapy performed by the administration to a subject of an expressed or expressible nucleic acid. In this embodiment of the invention, the nucleic acids produce their encoded antibody or prophylactic or therapeutic agent of the invention that mediates a prophylactic or therapeutic effect. Any of the methods for gene therapy available in the art can be used according to the present invention.

For general reviews of the methods of gene therapy, see Goldspiel et al. Methods commonly known in the art of recombinant DNA technology which can be used are described in Ausubel et al. Detailed description of various methods of gene therapy are disclosed in US A1 which is incorporated herein by reference.

RAGE plays a critical role in the pathology associated with a variety of diseases as defined herein above. Nature med. Using double-transgenic mice where the human APP gene is expressed and RAGE is overexpressed it was shown that overexpression of the normal RAGE gene leads to impairment in learning, increase in plaques whereas overexpression of a dominant-negative signalling defective RAGE variant leads to improvement in learning and lower plaque levels Arancio et al.

Experimentation in animal models of both Type 1 and 2 diabetes reveals that antagonism of the ligand-RAGE axis suppresses the development and progression of vascular and inflammatory cell perturbation in the diabetic milieu, e. Diabetes , 55 9 , ; De-Vriese et al.

Renal effects of a neutralising RAGE-antibody in long-term streptozotocin-diabetic mice. The Journal of endocrinology, , , Positive long-term renal effects of a neutralizing RAGE antibody in obese type 2 diabetic mice were shown by Flyvbjerg et al Diabetes, , 53, 1, p. In tumor-bearing mice, survival rates were prolonged, and spontaneous pulmonary metastases were inhibited by treatment using anti-RAGE neutralizing antibodies.

The antibodies, and antibody portions of the invention can be used to treat humans suffering from such a diseases. It should be understood that the antibodies of the invention or antigen binding portion thereof can be used alone or in combination with an additional agent, e. It should further be understood that the combinations which are to be included within this invention are those combinations useful for their intended purpose. The agents set forth below are illustrative for purposes and not intended to be limited.

The combinations, which are part of this invention, can be the antibodies of the present invention and at least one additional agent selected from the lists below. The combination can also include more than one additional agent, e. Non-limiting examples of therapeutic agents for multiple sclerosis with which an antibody, or antibody portion, of the invention can be combined include the following: It should be understood that the binding proteins and antibodies of the invention can be used alone or in combination with at least one additional agents suitable for treating one of the above diseases.

Said at least one additional agent may be selected by the skilled artisan for its intended purpose. For example, the additional agent can be a therapeutic agent such as a cholesterinase inhibitor e. The pharmaceutical compositions of the invention may include a "therapeutically effective amount" or a "prophylactically effective amount" of an antibody or antibody portion of the invention.

A "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of the antibody or antibody portion may be determined by a person skilled in the art and may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody or antibody portion to elicit a desired response in the individual.

A therapeutically effective amount is also one in which any toxic or detrimental effects of the antibody, or antibody portion, are outweighed by the therapeutically beneficial effects.

A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount.

Dosage regimens may be adjusted to provide the optimum desired response e. For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation.

It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

The specification for the dosage unit forms of the invention are dictated by and directly dependent on a the unique characteristics of the active compound and the particular therapeutic or prophylactic effect to be achieved, and b the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals. An exemplary, non-limiting range for a therapeutically or prophylactically effective amount of an antibody or antibody portion of the invention is 0.

It is to be noted that dosage values may vary with the type and severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that dosage ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition.

It will be readily apparent to those skilled in the art that other suitable modifications and adaptations of the methods of the invention described herein are obvious and may be made using suitable equivalents without departing from the scope of the invention or the embodiments disclosed herein.

Having now described the present invention in detail, the same will be more clearly understood by reference to the following examples, which are included for purposes of illustration only and are not intended to be limiting of the invention. Spleens from immunized animals were removed and single cell suspensions were prepared.

Spleen cells and tumor cells were mixed at a ratio of 5: Fused cells were seeded in 96 well plates in selective media, at a density of 2. Hybridomas that were producing mAbs with desired characteristics were subcloned by the limiting dilution method.

Colony PCR was performed on the transformants to identify clones containing insert. Inserts in the plasmids were sequenced on both strands to determine the variable heavy or variable light chain DNA sequences using M13 forward and M13 reverse primers Fermentas Life Sciences, Hanover MD. Variable heavy and variable light chain sequences of the 3 monoclonal antibodies 7F9, 11E6 and 4E5 and their three variable heavy chain CDRs and three variable light chain CDRs are listed in Table 4, above.

The light chain constant region of each of these antibodies was replaced by a human kappa constant region Table 1, above. Cell supernatants containing recombinant chimeric antibody were purified by Protein A Sepharose chromatography and bound antibody was eluted by addition of acid buffer.

Antibodies were neutralized and dialyzed into PBS. The reaction was stopped by addition of IN sulphuric acid. Plates were read spectrophotometrically at a wavelength of nm. Results are shown in Figures 1A, 1B , and 1C. The expression vector used for the generation of the stable expression was "pcDNA3 - 6.

Molecular biological standard techniques were used according to Sambrook and Russel Molecular Cloning: After transformation into E. The resulting plasmid "pcDNA 3 - 6.

For expression and purification of sufficient amounts of husRAGE protein this clone was grown in serum containing cell culture medium see above in cell factories Nunc, Germany. Equilibration of the column and binding of the hexa-His containing protein from cell supernatants to the matrix were done according to instructions by the manufacturer.

Elution of the protein was done using step gradients with increasing concentrations of imidazole. Positive fractions were combined, concentrated and dialyzed 3 times against PBS 2 x 4h, 1 x 16 h. This protein was generated by the same basic procedure used for husRAGE After agarose gel and elution of the fragment the resulting pure fragment was cleaved with restriction endonucleases HindIII and XhoI and purified again using agarose gel and elution.

Equilibration and binding were done according to instruction by the manufacturer. Purified husRAGE concentrations were determined spectrophotometrically. After agarose gel and elution of the fragment the resulting pure fragment was cleaved with restriction endonucleases NcoI and XhoI and purified again using agarose gel and elution. The ligation mixtures were transformed into E. A positive clone was picked and the plasmid DNA isolated. Expression of this plasmid using the Freestyle Expression system and F cells Example 2.

Positive clones were identified and the resulting plasmid DNA purified named: The resulting plasmid was named: HEK F cells that had been grown in culture for 2 - 3 days in Free Style Expression Medium were centrifuged at g and the supernatant discarded.

Transfection mixtures with fectin-DNA complex were set up as follows:. DNA mixture and fectin-solution from i and ii were transferred to a new tube, mixed slightly and after incubation for 25 minutes at room temperature were added to the cells in the Erlenmeyer. Cell supernatants were harvested by centrifugation at g for 10 minutes. To couple the protein from cell supernatants to beads, beads protein G-sepharose 4 Fast Flow Amersham Bioscience were washed 3 times in PBS by suspending the beads in PBS and centrifugation at 13, rpm, discarding the supernatant.

Beads were incubated with the respective cell supernatants ml cell supernatants per ml beads to be coupled for 1 - 2 hours on a rotator at room temperature. After incubation the beads were washed 3 times with PBS as above. After centrifugation the supernatant was immediately neutralized by adding 2 M Tris to adjust the pH to 7. The bead pellet was discarded. Dot blots were used to evaluate the binding of antibodies to peptides or fragments of RAGE in a non-denatured form.

The identities of peptides were verified by mass spectrometry. Peptides used were 30mers spanning the extracellular region of the human RAGE protein. Net charge of most peptides was similar. Membranes were dried and unspecific binding was blocked by shaking the membranes for 1 hour at room temperature with Western Blocking reagent Roche, no. Get to Know Us. Amazon Payment Products. English Choose a language for shopping.

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Rothenberg, D. Munro, et al. Lawrence Livermore National Laboratory, September 24, Science and Technology Review. Plasmas 5, , , doi: McCrory, S. Skupsky, et al. Albritton, and E. When Peer Review Fails Technical report. New York Times. The Scientist. T et all, Target area for Nova Upgrade: CS1 maint: Powell and Richard H.

Archived from the original PDF on Fusion Fun: Innovation America. August 30, October 11, BBC News. Technology Review. Scientific American. May Project Status — May. Retrieved 21 Mar March 22, Princeton Plasma Physics Laboratory. Broad September 30, The New York Times. Ignition switch". Physics Today. British Broadcasting Corporation. Uh, Not Really". Contra Cost Times. San Jose Mercury News. The Independent. Tschudi, Jannick P.


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