In vitro Evaluation of Skin Related Enzyme Inhibitory Effect and Emulgel Formulation Development Studies of Onobrychis Argyrea subsp. Argyrea with Phytochemical Analysis.

Species of Onobrychis have been used to treat skin disorders such as wounds and cuts in folk medicine and Onobrychis argyrea subsp. argyrea (OA) commonly known as 'silvery sainfoin', is a member of this genus. In this study, it was aimed to investigate the skin-related biological activities and phytochemical characterization of OA. Moreover, an emulgel formulation was developed from the main methanolic extract of the plant (OAM). Initially, to identifiy of the active fractions, aerial parts of the plant material was extracted with methanol and fractionated by n-hexane, chloroform, ethyl acetate and n-butanol, respectively. Antioxidant activity was determined by CUPRAC, TOAC, FRAP and DPPH assays. Thereafter, the inhibition potential of OAM, novel formulation and all fractions was measured against elastase, collagenase, tyrosinase and hyaluronidase enzymes. OAM was analyzed and characterized by LC/MS-MS. The major bioactive flavonoids which are rutin and isoquercetin were measured and compared as qualitative and quantitative via high performance thin layer chromatography (HPTLC) analysis in OAM and fractions. The results showed that extracts of OA can be a potential cosmeceutical agent for skin related problems.

Microwave-assisted rapid conjugation of horseradish peroxidase-dextran aldehyde with Schiff base reaction and decolorization of Reactive Blue 19.

Microwave irradiation has become a routine technique in homogeneous and effective heating in organic synthesis. However, its application in enzyme-containing reactions is limited since it can cause denaturation of the enzyme. In this study, we have briefly investigated the effect of microwave heating on the conjugation reaction of horseradish peroxidase (HRP) with aldehyde derivative of dextran (D-CHO). The reaction was irradiated by microwave at 50 °C for 5 min. The conjugate was confirmed via GPC, in which the conjugates of HRP and D-CHO coexist with free unbound HRP molecules. Activity studies of HRP revealed that there is a small decrease in conjugate activity relative to the free enzyme after a short bioconjugation reaction with microwave irradiation. In decolorization studies of the textile dye Reactive Blue 19 (RB19), 99% of RB19 was decolorized through the free enzyme at 35 °C while the decolorization of the dye was 96% at 25-35 °C by the conjugate, which is a critical result showing clearly that the HRP conjugated via D-CHO is not denatured and still active after microwave-assisted reaction. This phenomenon is due to the multiple point conjugation of D-CHO on the surface of HRP and locking the 3D structure which may prevent changes in the secondary or tertiary structure of the enzyme. The results reveal that microwave irradiation can be used in production of covalently modified enzymes.

Determination of minimum enzymatic decolorization time of reactive dye solution by spectroscopic & mathematical approach.

Synthetic dyes are very important for textile dyeing, paper printing, color photography and petroleum products. Traditional methods of dye removal include biodegradation, precipitation, adsorption, chemical degradation, photo degradation, and chemical coagulation. Dye decolorization with enzymatic reaction is an important issue for several research field (chemistry, environment) In this study, minimum decolorization time of Remazol Brilliant Blue R dye with Horseradish peroxidase enzyme was calculated using with mathematical equation depending on experimental data. Dye decolorization was determined by monitoring the absorbance decrease at the specific maximum wavelength for dye. All experiments were carried out with different initial dye concentrations of Remazol Brilliant Blue R at 25 degrees C constant temperature for 30 minutes. The development of the least squares estimators for a nonlinear model brings about complications not encountered in the case of the linear model. Decolorization times for completely removal of dye were calculated according to equation. It was shown that mathematical equation was conformed exponential curve for dye degradation.

Inhibition of mammalian carbonic anhydrase isoforms I, II and VI with thiamine and thiamine-like molecules.

Here we determined the in vitro inhibitory effects of 5-(2-hydroxyethyl)-3,4-dimethylthiazolium iodide (1), 3-Benzyl-5-(2-hydroxyethyl)-4-methylthiazolium chloride (2) and thiamine (3) on human erythrocyte carbonic anhydrase I, II isozymes (hCA I and hCA II) and secreted isoenzyme CA VI. K(I) values ranged from 0.38 to 2.27 µM for hCA I, 0.085 to 0.784 µM for hCA II and 0.062 to 0.593 µM for hCA VI, respectively. The compounds displayed relatively strong actions on hCA II, in the same range as the clinically used sulfonamidesethoxzolamide, zonisamide and acetazolamide.