An Extreme Vertices Mixture Design Approach to Optimization of Tyrosinase Inhibition Effects

The objective of this study is to optimize the tyrosinase inhibition effects of three mixtures included Emblica extract, L-Ascorbic acid, and Kojic acid. Tyrosinase is a copper-containing oxidase, which has activity for both catechol and cresol. It is responsible for browning reactions. Mushroom tyrosinase was used in this study as the tyrosinase source and L-DOPA was used to activate the browning activity. The seventeen formulations were conducted by extreme vertices mixture design. To specify the lower and upper limit, prior study for each ingredient was experimented. Ten and thirty-three percentage of inhibition were proposed to specify the lower and upper limit of each ingredient. The result of experiment shown the difference tyrosinase inhibition effects of seventeen formulations. The analysis of variance for result was significant for all pairs and all of components with 92.89 percentage of R-square adjusted. To optimize the best combination, used of response optimization by setting up the target to 80, 85, 90, and 95 percentage of tyrosinase inhibition effect. The optimum combination with 90%inhibition target is 3.1541 mg./ml. of Emblica extract, 0.1420 mg./ml. of L-Ascorbic acid, and 0.0297 mg./ml. of Kojic acid, this formula unlike the formula that was experimented on.The objective of this study is to optimize the tyrosinase inhibition effects of three mixtures included Emblica extract, L-Ascorbic acid, and Kojic acid. Tyrosinase is a copper-containing oxidase, which has activity for both catechol and cresol. It is responsible for browning reactions. Mushroom tyrosinase was used in this study as the tyrosinase source and L-DOPA was used to activate the browning activity. The seventeen formulations were conducted by extreme vertices mixture design. To specify the lower and upper limit, prior study for each ingredient was experimented. Ten and thirty-three percentage of inhibition were proposed to specify the lower and upper limit of each ingredient. The result of experiment shown the difference tyrosinase inhibition effects of seventeen formulations. The analysis of variance for result was significant for all pairs and all of components with 92.89 percentage of R-square adjusted. To optimize the best combination, used of response optimization by setting up the target to 80, 85, 90, and 95 percentage of tyrosinase inhibition effect. The optimum combination with 90% inhibition target is 3.1541 mg/ml of Emblica extract, 0.1420 mg/ml of L-Ascorbic acid, and 0.0297 mg/ml of Kojic acid, this formula unlike the formula that was experimented on

Gamma Irradiation Causes Variation and Stability of Artemisinin Content in <em>Artemisia annua</em> Plants

Artemisinin is an anti-malarial sesquiterpene lactone isolated from Artemisia annua L., a traditional Chinese herb of the family Asteraceae. The plant contains relatively low artemisinin content, ranging from 0.01 to 0.8% of the plant dry weight, depending on the geographical origin, seasonal, and somatic variations. Ionizing radiation has been recognized as a powerful technique for plant improvement, especially in crop plants. This technique creates genetic variability in plants, which can be screened for desirable characteristics. Very little is known about the effect of gamma irradiation on the potential increase of artemisinin production in A. annua. In this study, 130 shoot tips excised from the population of in vitro A. annua plantlets (with an average leaf artemisinin content of 0.18 ± 0.09%) were exposed to 5 Gy 60Co gamma irradiation and subsequently transferred to a suitable medium for in vitro development of plantlets. The resulting 90 stable survived after four passages appeared to have a wide variation of artemisinin content, ranging from 0.02 to 0.68% of dry weight. All the viable plantlets were then transferred from the in vitro cultures to ex vitro conditions both in a greenhouse and an open field. A significant correlation was observed between artemisinin content among individual pairs of the vitro plantlets and ex vitro mature plants, with the correlation coefficient (R2) values of 0.915 for the greenhouse plants and 0.797 for the open field plants. Among these, the highest artemisinin-containing plant appeared to accumulate 0.84% artemisinin of dry weight in the open field, which is almost five times higher than the original plants. These results suggest that gamma irradiation with 5-Gy dose can produce viable variants of A. annua that can maintain the biosynthetic capability of artemisinin throughout the in vitro-ex vitro transfer and development of the first generation of mature plants

Arthrogenicity of type II collagen monoclonal antibodies associated with complement activation and antigen affinity

<p>Abstract</p> <p>Background</p> <p>The collagen antibody-induced arthritis (CAIA) model, which employs a cocktail of monoclonal antibodies (mAbs) to type II collagen (CII), has been widely used for studying the pathogenesis of autoimmune arthritis. In this model, not all mAbs to CII are capable of inducing arthritis because one of the initial events is the formation of collagen-antibody immune complexes on the cartilage surface or in the synovium, and subsequent activation of the complement by the complexes induces arthritis, suggesting that a combination of mAbs showing strong ability to bind mouse CII and activate the complement may effectively induce arthritis in mice. In the present study, we examined the relationship between the induction of arthritis by the combination of IgG2a (CII-6 and C2A-12), IgG2b (CII-3, C2B-14 and C2B-16) and IgM (CM-5) subclones of monoclonal antibodies (mAb) of anti-bovine or chicken CII and the ability of mAbs to activate complement and bind mouse CII.</p> <p>Methods</p> <p>DBA/1J mice were injected with several combinations of mAbs followed by lipopolysaccharide. Furthermore, the ability of mAbs to activate the complement and bind mouse CII was examined by ELISA.</p> <p>Results</p> <p>First, DBA/1J mice were injected with the combined 4 mAbs (CII-3, CII-6, C2B-14, and CM-5) followed by lipopolysaccharide, resulting in moderate arthritis. Excluding one of the mAbs, i.e., using only CII-3, CII-6, and C2B-14, induced greater inflammation of the joints. Next, adding C2A-12 but not C2B-16 to these 3 mAbs produced more severe arthritis. A combination of five clones, consisting of all 5 mAbs, was less effective. Histologically, mice given the newly developed 4-clone cocktail had marked proliferation of synovial tissues, massive infiltration by inflammatory cells, and severe destruction of cartilage and bone. Furthermore, 4 of the 6 clones (CII-3, CII-6, C2B-14, and C2A-12) showed not only a strong cross-reaction with mouse CII but also marked activation of the complement <it>in vitro</it>.</p> <p>Conclusion</p> <p>The combination of 4 mAbs showing strong abilities to activate the complement and bind mouse CII effectively induced arthritis in DBA/1J mice. This <it>in vitro </it>system may be useful for the selection of mAbs associated with the development of arthritis.</p