Abstract: Novel enzymatic methods to determine the concentration of homocysteine in biological fluids are described. In a typical embodiment of the invention, the biological fluid sample is from a patient, and the methods of the invention are useful to assess risk for cardiovascular disease. The novel methods of the invention involve use of particular homocysteinase enzymes that permit the determination of homocysteine concentrations in biological samples without interference from the concentrations of cysteine and/or of methionine that are routinely present in such samples. There is also provided a diagnostic kit for use in determining the amount of homocysteine in a biological sample comprising (a) a homocysteinase having the aforementioned characteristics, and (b) at least one reagent capable of being used to determine the amount of product formed in the homocysteinase reaction.

Abstract: Novel enzymatic methods to determine the concentration of homocysteine in biological fluids are described. In a typical embodiment of the invention, the biological fluid sample is from a patient, and the methods of the invention are useful to assess risk for cardiovascular disease. The novel methods of the invention involve use of particular homocysteinase enzymes that permit the determination of homocysteine concentrations in biological samples without interference from the concentrations of cysteine and/or of methionine that are routinely present in such samples. There is also provided a diagnostic kit for use in determining the amount of homocysteine in a biological sample comprising (a) a homocysteinase having the aformentioned characteristics, and (b) at least one reagent capable of being used to determine the amount of product formed in the homocysteinase reaction.

Abstract: Novel enzymatic methods to determine the concentration of homocysteine in biological fluids are described. In a typical embodiment of the invention, the biological fluid sample is from a patient, and the methods of the invention are useful to assess risk for cardiovascular disease. The novel methods of the invention involve use of particular homocysteinase enzymes that permit the determination of homocysteine concentrations in biological samples without interference from the concentrations of cysteine and/or of methionine that are routinely present in such samples. There is also provided a diagnostic kit for use in determining the amount of homocysteine in a biological sample comprising (a) a homocysteinase having the aformentioned characteristics, and (b) at least one reagent capable of being used to determine the amount of product formed in the homocysteinase reaction.

Abstract: The present invention provides a method to specifically target hair follicles with formulations which effect the production of hair coloring pigments in the follicle. Liposomal formulations for this purpose are disclosed.

Abstract: The invention provides methods to deliver macromolecules to hair follicles selectively using formulations of these macromolecules in liposomal separations.

Abstract: The present invention discloses the cloning of a methioninase-encoding nucleic acid molecule from Pseudomonas putida and the construction of high-level expression modules containing the methioninase-encoding nucleic acid molecule. The invention further provides expression modules that use the T7 RNA polymerase promoter to express the isolated methioninase-encoding nucleic acid molecules. Expression modules employing the T7 promoter were found to produce unexpectedly high levels of methioninase. The present invention further provides purification methods to obtain highly pure, endotoxin free methioninase, chemically modified forms of methioninase, crystallized methioninase and lyophilized methioninase preparations. The present invention further provides therapeutical methods using the disclosed recombinant methioninase preparations.

Abstract: Improved formulations of methioninase which contain methioninase in crystalline, lyophilized, or conjugated form are disclosed. The methioninase in these compositions has a specific activity of at least 20 U/mg and the formulations are detergent free and contain less than 1 ng endotoxin per mg protein.

Abstract: Methods and compositions for a native-state histoculturing system for skin are disclosed. Skin samples are placed on an extracellular support matrix immersed in a medium whereby the internal surface of the skin is adjacent to the matrix and the external surface is exposed above the surface of the medium and the skin maintained in the medium under skin culturing conditions. An extracellular support matrix comprising a collagen-containing gel and homopolysaccharide sponge or a combination matrix of a top layer of a collagen-containing gel and a bottom layer of a homopolysaccharide sponge is also disclosed. Skin toxicity, hair growth, anti-aging and anti-wrinkling assays utilizing the histoculturing system of the present invention are also disclosed.

Abstract: The present invention provides a methods and apparatuses for use in providing rapid, selective and effective targeted delivery of beneficial substances into hair follicles. The invention relies on the use of vibrational method of delivery of substances of different size from small molecules up to particles compatible with the diameter of hair strand into the hair bulb. The skin is subjected to the vibration of about 1 Hz to about 100 Hz frequency and 0.1 mm to about 10 mm amplitude, an amount which is sufficient to cause mechanical shifts in different directions of skin structure without damage. This results in follicle effective uptake of the tested substances such as carbon particles of different size as well as melanin. The methods and apparatuses of the present invention are particularly useful in conjunction with photosensitive agents and electromagnetic irradiation. Such a combination is used to target the delivery of the agent to within hair follicles and then subsequently activate the agent.

Abstract: The invention describes a method to deliver a composition selectively to hair follicles using a liposomal formulation. Proteins which are cell cycle inhibitors are products of the multi-drug resistance gene or the recombinant materials for their production are targeted to hair follicles by encapsulating them in liposomes.

Abstract: The present invention relates to a method of using an in vitro culture system to measure the cellular proliferative capacity cellular viability of human tissues, particularlytumor tissues. The invention also describes the use of the method to evaluate the effectiveness of an antineoplastic drug in inhibiting tumor cell proliferation of viability.

Abstract: The present invention provides methods for treating and reducing the potential for cardiovascular disease using methioninase compositions having less than 10 ng endotoxin per mg methioninase.

Abstract: The present invention describes methods for using methioninase as an antitumor reagent in anti-methionine chemotherapy. Specifically, methioninase is used to slow or stop cell division which can be enhanced with competitive inhibitors of methionine. In addition, methioninase can be used in combination with chemotherapeutic agents to increase the therapeutic effectiveness of the agent by inducing cell cycle synchronization.

Abstract: The present invention describes a method for targeted and specific delivery of beneficial compounds, including dyes, proteins, and nucleic acids for gene therapy, to hair follicle cells using liposomes encapsulating the beneficial compound. Particularly preferred methods describe delivery of tyrosinase to the hair follicle for the purpose of improving hair color or condition, either by encapsulating the compound in liposomes, or by encapsulating a nucleic acid capable of expressing the protein in liposomes.

Abstract: A method of evaluating the effectiveness of drugs in inhibiting the growth of tumor cells. A sample of a tumor is histocultured, a drug to be evaluated is added to the sample and the sample is incubated. A suitable tetrazolium salt is added and a frozen section of the sample is prepared. The section is stained with a fluorescent dye. The section is exposed to polarized light and the reflected light is measured. Then the section is exposed to fluorescent light and the light emitted by the dye is measured. These measurements are compared to a control and/or measurements from tests using other drugs and the relative effectiveness of the drug is evaluated, preferably by pixel analysis.