Analysis of transition state mimicry by tight binding aminothiazoline inhibitors provides insight into catalysis by human O-GlcNAcase.

The modification of nucleocytoplasmic proteins with O-linked N-acetylglucosamine (O-GlcNAc) plays diverse roles in multicellular organisms. Inhibitors of O-GlcNAc hydrolase (OGA), the enzyme that removes O-GlcNAc from proteins, lead to increased O-GlcNAc levels in cells and are seeing widespread adoption in the field as a research tool used in cells and in vivo. Here we synthesize and study a series of tight binding carbohydrate-based inhibitors of human OGA (hOGA). The most potent of these 2'-aminothiazolines binds with a sub-nanomolar Ki value to hOGA (510 ± 50 pM) and the most selective has greater than 1 800 000-fold selectivity for hOGA over mechanistically related human lysosomal ß-hexosaminidase. Structural data of inhibitors in complex with an hOGA homologue reveals the basis for variation in binding among these compounds. Using linear free energy analyses, we show binding of these 2'-aminothiazoline inhibitors depends on the pKa of the aminothiazoline ring system, revealing the protonation state of the inhibitor is a key driver of binding. Using series of inhibitors and synthetic substrates, we show that 2'-aminothiazoline inhibitors are transition state analogues of hOGA that bind to the enzyme up to 1-million fold more tightly than the substrate. These collective data support an oxazoline, rather than a protonated oxazolinium ion, intermediate being formed along the reaction pathway. Inhibitors from this series will prove generally useful tools for the study of O-GlcNAc. The new insights gained here, into the catalytic mechanism of hOGA and the fundamental drivers of potency and selectivity of OGA inhibitors, should enable tuning of hOGA inhibitors with desirable properties.

A combined approach in characterization of an effective w/o hand cream: the influence of emollient on textural, sensorial and in vivo skin performance.

To formulate a consumer-acceptable cosmetic product, numerous demands have to be fulfilled, and as the most important, efficacy (both real and perceived), adequate aesthetic (visual perception) and all sensorial characteristics have to be achieved. In this study, four model water-in-oil creams intended for hand care, varying in one emollient component, were submitted to rheological, sensory and textural characterization, and their efficacy was evaluated in in vivo study on human volunteers. Our results indicate that certain alteration restricted to the oil phase induced a change in all investigated characteristics, showing that each instrumental measurement can be used as a sensitive tool in the characterization of cream samples. Regarding the correlation between physical measurements and certain sensory attributes, it is possible to formulate a product with specific sensory characteristics by using pre-defined rheological or textural parameters. To obtain a complete sensory profile of a cosmetic product, a detailed sensory evaluation should be carried out according to the existing standard practices, which are both time- and money-consuming. However, a modified sensory study could be useful for fast in-line screening along with instrumental characterization of a novel cosmetic emulsion product and could be particularly helpful in the process of distinguishing a single formulation from several differing in one component.

Vehicle-controlled effect of urea on normal and SLS-irritated skin.

It is known that, depending on the concentration, treatment with urea could improve skin barrier function, despite its penetration-enhancing properties. This controversial skin effect of urea has been explored systematically in this study in terms of the effect of vehicle on the performance of urea. In the first part, a series of four semi-solid emulsions with 5% (w/w) urea, varying in the type of emulsion, nature of emulsifier and polarity of oil ingredients, have been evaluated with regard to their skin hydrating and transepidermal water loss (TEWL)-modifying properties. Placebo samples were tested alongside the urea-containing ones. Two best performing moisturisers from the above were chosen for the second part of the study, in which sodium lauryl sulphate (SLS)-irritated skin was treated with both placebo and urea-containing samples. In addition to TEWL and skin hydration level, the erythema index (EI) was measured before, during and after the treatment. The results have shown that barrier-improving and hydrating abilities of urea are bi-directional and dependent on both the type of vehicle used for its delivery and the state of skin.