Abstract: Compositions comprising ergothioneine and a trimethylamine absorber are provided. Also provided are methods for preventing, reducing or minimizing the fishy, amine odor, due to trimethylamine, that is associated with the processing and/or storage of a preparation containing ergothioneine, by combining with the ergothioneine, during processing or prior to storage, a trimethylamine absorber in an amount sufficient to prevent the detection of any trimethylamine odor by the human nose. A method is further provided for ameliorating the methylamine odor associated with an aqueous ergothioneine-containing preparation after it has developed a fishy trimethylamine odor.

Abstract: Recombinant forms of DNA sequences for CPD glycosylases, including the bacteriophage T4 gene denV, are described that are capable of expression at high levels. Active CPD glycosylases can be recovered from inclusion bodies resulting from the high expression using, for example, a homogenization process which employs stream mixing, and the active proteins can be used in, for example, topical formulations for treatment of photosensitive diseases. Stream mixing can also be used to solubilize inclusion bodies containing proteins other than CPD glycosylases.

Abstract: Recombinant forms of DNA sequences for CPD glycosylases, including the bacteriophage T4 gene denV, are described that are capable of expression at high levels. Active CPD glycosylases can be recovered from inclusion bodies resulting from the high expression using, for example, a homogenization process which employs stream mixing, and the active proteins can be used in, for example, topical formulations for treatment of photosensitive diseases. Stream mixing can also be used to solubilize inclusion bodies containing proteins other than CPD glycosylases.

Abstract: Compositions and methods are provided for reversing and/or inhibiting inflammation, e.g., by inhibiting prostaglandin and/or COX-2 production, using one or more indolequinazoline alkaloids, preferably in combination with butylated hydroxytoluene. The preferred indolequinazoline alkaloids are rutaecarpine, evodiamine, and dehydroevodiamine, which are naturally found in unpurified form in the traditional Chinese medicine Wu Chu Yu made from the fruit of the herb, Evodia rutaecarpa.

Abstract: The invention provides T4 endonuclease V compositions that exhibit enhanced stability, including stability at non-refrigerated temperatures, through reduced activity of cryptic proteases. Methods for detecting cryptic protease activity and methods for reducing such activity are also provided.

Abstract: The topical use of ursolic acid compounds to alter the lipid content of mammalian skin is disclosed. The compounds can be encapsulated in liposomes and administered in this form to the skin in, for example, a lotion or a gel. The compounds are effective in, among other things, reducing the effects of aging, photoaging, and skin atrophy, including skin atrophy resulting from the topical use of retinoids and/or steroids. Compositions comprising a ursolic acid compound in combination with another therapeutically active topical compounds, such as, a retinoid or a steroid, are also disclosed.

Abstract: The invention provides T4 endonuclease V compositions that exhibit enhanced stability, including stability at non-refrgerated temperatures, through reduced activity of cryptic proteases. Methods for detecting cryptic protease activity and methods for reducing such activity are also provided.

Abstract: Protection of mitochondria from oxidative damage due to natural or disease processes as well as by the effects of exogenous factors such as incident sunlight, exposure via inhalation to oxidative environmental toxins, consumption of dietary oxidants, and oxidative-stress-inducing cosmetics and pharmaceuticals, radiation therapy, among others, is provided by a composition comprising L-ergothioneine, L-ergothioneine may be prepared in a cosmetically or pharmaceutically-acceptable base to form an agent for topical application to the skin, and for oral or parenteral administration. Effective application and delivery, of L-ergothioneine is enhanced by encapsulation in a liposomes, a preferred embodiment. Diagnostic methods for determining exposure and susceptibility to radiation, radical and reactive oxygen species in mammals is also provided.

Abstract: Protection of mitochondria from oxidative damage due to natural or disease processes as well as by the effects of exogenous factors such as incident sunlight, exposure via inhalation to oxidative environmental toxins, consumption of dietary oxidants, and oxidative-stress-inducing pharmaceuticals, exposure to radiation including radiation therapy, among others, is provided by a composition comprising L-ergothioneine. L-ergothioneine may be prepared in a pharmaceutically-acceptable carrier to form an agent for topical application to the skin, and for orally or parenteral administration. Effective application and delivery of L-ergothioneine is enhanced by encapsulation in a liposome, a preferred embodiment. Diagnostic methods for determining exposure and susceptibility to radiation, radical, and reactive oxygen species in mammals is also provided.

Abstract: Assays for the DNA repair protein O.sup.6 -methylguanine-DNA methyltransferase (MGMT) are provided which employ monoclonal antibodies prepared using MGMT having transferase activity, as opposed to denatured MGMT or MGMT fragments. The monoclonal antibodies are able to recognize MGMT in single cell preparations (immunohistochemical staining assays) and in cell extracts (immunoassays). In connection with immunohistochemical staining, the use of a fluorescent readout coupled with digitization of the cell image allows for quantitative measures of MGMT levels in, for example, tumor biopsy samples. Such quantitative measures can be used to determine which patients are likely to benefit from chemotherapy using alkylating agents since tumor cells having low MGMT levels are more likely to be killed by such agents than those with high MGMT levels.

Abstract: A method for tanning skin is provided in which liposomes containing a DNA repair enzyme are administered to skin in combination with exposure of the skin to UV radiation. The result is an enhanced level of melanin production, i.e., more tanning than achieved by UV radiation alone. The administration of the DNA repair enzymes in liposomes also reduces the level of DNA damage caused by the UV exposure. Accordingly, both the tanning response is increased and the deleterious effect of UV exposure is decreased. The method can be used by the general population as well as by individuals whose skin is susceptible to UV-induced damage.

Abstract: Exposing the skin to UV radiation interferes with the induction of the T-cell mediated immune response, including both delayed (DHS) and contact (CHS) hyper-sensitivity immune responses initiated at non-irradiated sites. The present inventors have discovered that DNA is at least one of the targets for UV-induced hypersensitivity, and demonstrate that the application of DNA repair enzymes can reverse the damaging effects of UV irradiation on both the DHS and CHS response. The usefulness of the invention in this regard was tested using a model immunosuppression system in mice. In these studies, mice were first exposed to UV radiation and then liposomes were used to deliver a dimer-specific excision repair enzyme to their epidermis in situ. The application of liposomal T4 endonuclease V encapsulated to the UV-irradiated skin both decreased the number of cyclobutane pyrimidine dimers in the epidermis and prevented suppression of both delayed and contact hypersensitivity responses.

Abstract: Methods for purifying DNA repair enzymes are provided in which an aqueous solution of a DNA repair enzyme in an impure state is applied to a molecular sieve column having an exclusion limit which will retard the DNA repair enzyme but will not retard contaminants larger than the DNA repair enzyme. The DNA repair enzyme in an enhanced state of purity is eluted isocratically from the molecular sieve column in an elution buffer and applied directly to a DNA affinity column in the same buffer without intermediate dialysis, ultrafiltration, or other procedures. The DNA repair enzyme is eluted from the DNA affinity column using, for example, a salt gradient. The method is rapid, inexpensive, simple to perform, and has been found to produce a homogeneous final product. In accordance with other aspects of the invention, the purified DNA repair enzymes are encapsulated in liposomes and administered to living cells in situ.

Abstract: Methods for purifying DNA repair enzymes are provided in which an aqueous solution of a DNA repair enzyme in an impure state is applied to a molecular sieve column having an exclusion limit which will retard the DNA repair enzyme but will not retard contaminants larger than the DNA repair enzyme. The DNA repair enzyme in an enhanced state of purity is eluted isocratically from the molecular sieve column in an elution buffer and applied directly to a DNA affinity column in the same buffer without intermediate dialysis, ultrafiltration, or other procedures. The DNA repair enzyme is eluted from the DNA affinity column using, for example, a salt gradient. The method is rapid, inexpensive, simple to perform, and has been found to produce a homogeneous final product. In accordance with other aspects of the invention, the purified DNA repair enzymes are encapsulated in liposomes and administered to living cells in situ.

Abstract: A method for administering a protein having intracellular biological activity into the interior of living skin cells, which lie below the skin's stratum corneum, is provided. The method comprises the steps of: (a) encapsulating the protein in liposomes; and (b) applying the liposomes to the outer surface of living skin so that the protein encapsulated in the liposomes traverses the skin's stratum corneum and the outer membranes of said cells and is thereby delivered by the liposomes into the interior of said cells. In certain preferred embodiments, the liposomes are pH sensitive liposomes. In other preferred embodiments, the protein is a DNA repair enzyme, such as T4 endonuclease V.

Abstract: Methods for purifying DNA repair enzymes are provided in which an aqueous solution of a DNA repair enzyme in an impure state is applied to a molecular sieve column having an exclusion limit which will retard the DNA repair enzyme but will not retard contaminants larger than the DNA repair enzyme. The DNA repair enzyme in an enhanced state of purity is eluted isocratically from the molecular sieve column in an elution buffer and applied directly to a DNA affinity column in the same buffer without intermediate dialysis, ultrafiltration, or other procedures. The DNA repair enzyme is eluted from the DNA affinity column using, for example, a salt gradient. The method is rapid, inexpensive, simple to perform, and has been found to produce a homogeneous final product. In accordance with other aspects of the invention, the purified DNA repair enzymes are encapsulated in liposomes and administered to living cells in situ.