Hemisynthesis of Anthocyanin Phase II Metabolites by Porcine Liver Enzymes.

The aim of this work was to obtain phase II metabolites of cyanidin-3- O-glucoside and its aglycone using porcine liver enzymes. For this purpose, anthocyanins extracted from blackberry concentrate and containing mostly cyanidin-3- O-glucoside were incubated with the S9, microsomal, and cytosolic fractions of porcine liver. The reactions were targeted to the direction of the respective phase II transformation by the addition of activated cofactors. LC-MS n and LC-IMS-QTOF-MS analyses showed that one methylated, three glucuronidated and three sulfated metabolites of cyanidin-3- O-glucoside were generated. The aglycone, cyanidin, was sulfated and glucuronidated by the liver enzymes. In addition, both were glucuronidated and methylated simultaneously. The detected compounds and the generated data like exact masses, mass spectra, and CCS values may serve as a basis in the search for metabolites formed in vivo. As their effects are largely unexplored, the described synthesis may contribute to a better understanding of the metabolism of anthocyanins.

Use of complementary and alternative medicine in healthy children and children with chronic medical conditions in Germany.

OBJECTIVES: Use of complementary and alternative medicine (CAM) in children is common and probably increasing. However little is known about differences between healthy and chronically ill children with a focus on prevalence, reasons for use/non-use, costs, adverse effects and socio-demographic factors. DESIGN: A questionnaire-based survey with 500 participants visiting the outpatient clinic of the University Children's Hospital Homburg, Germany was conducted over a 4-week period in 2004. Recruitment was stopped when 500 questionnaires were handed out in total. RESULTS: Of the 405 (81%) respondents (242 with chronic conditions, 163 healthy children incidentally visiting the hospital for minor ailments) 229 (57%) reported lifetime CAM use (59% with chronic conditions versus 53% healthy children). Among CAM users the most prevalent therapies were homeopathy (25%), herbal remedies (8%), anthroposophic medicine (7%), vitamin preparations (6%) and acupuncture (5%). The main reasons for use were to strengthen the immune system, physical stabilisation and to increase healing chances/maintain health. Socio-demographic factors associated with CAM use were tertiary education (mother: p=0.017; father: p>0.001), higher family income (p=0.001) and being Protestant (p=0.01). Expectations towards CAM were high and most parents would recommend certain CAM (94%). 79% of the users informed a physician about CAM use. Side effects were rarely reported (4%), minor and self-limiting. CONCLUSIONS: Clinical care and the physician-patient relation would benefit from an enhanced understanding of CAM and a greater candidness towards the parental needs. The safety and efficacy especially of CAM with high prevalence rates should be determined in rigorous basic and clinical researches.

Stretching mechanotransduction from the lung to the lab: approaches and physiological relevance in drug discovery.

Recent years have shown a great deal of interest and research into the understanding of the biological and physiological roles of mechanical forces on cellular behavior. Despite these reports, in vitro screening of new molecular entities for lung ailments is still performed in static cell culture models. Failure to incorporate the effects of mechanical forces during early stages of screening could significantly reduce the success rate of drug candidates in the highly expensive clinical phases of the drug discovery pipeline. The objective of this review is to expand our current understanding of lung mechanotransduction and extend its applicability to cellular physiology and new drug screening paradigms. This review covers early in vivo studies and the importance of mechanical forces in normal lung development, use of different types of bioreactors that simulate in vivo movements in a controlled in vitro cell culture environment, and recent research using dynamic cell culture models. The cells in lungs are subjected to constant stretching (mechanical forces) in regular cycles due to involuntary expansion and contraction during respiration. The effects of stretch on normal and abnormal (disease) lung cells under pathological conditions are discussed. The potential benefits of extending dynamic cell culture models (screening in the presence of forces) and the associated challenges are also discussed in this review. Based on this review, the authors advocate the development of dynamic high throughput screening models that could facilitate the rapid translation of in vitro biology to animal models and clinical efficacy. These concepts are translatable to cardiovascular, digestive, and musculoskeletal tissues and in vitro cell systems employed routinely in drug-screening applications.