Two parametric cell cycle analyses of plant cell suspension cultures with fragile, isolated nuclei to investigate heterogeneity in growth of batch cultivations.

Plant cell suspensions are frequently considered to be heterogeneous with respect to growth in terms of progression of the cells through the cell cycle and biomass accumulation. Thus, segregated data of fractions in different cycle phases during cultivation is needed to develop robust production processes. Bromodeoxyuridine (BrdU) incorporation and BrdU-antibodies or 5-ethynyl-2'-deoxyuridine (EdU) click-it chemistry are frequently used to acquire such information. However, their use requires centrifugation steps that cannot be readily applied to sensitive cells, particularly if nuclei have to be extracted from the protective cellular milieu and envelopes for DNA analysis. Therefore, we have established a BrdU-Hoechst stain quenching protocol for analyzing nuclei directly isolated from delicate plant cell suspension cultures. After adding BrdU to test Harpagophytum procumbens cell suspension cultures the cell cycle distribution could be adequately resolved using its incorporation for the following 72 h (after which BrdU slowed biomass accumulation). Despite this limitation, the protocol allows resolution of the cell cycle distribution of cultures that cannot be analyzed using commonly applied methods due to the cells' fragility. The presented protocol enabled analysis of cycling heterogeneities in H. procumbens batch cultivations, and thus should facilitate process control of secondary metabolite production from fragile plant in vitro cultures. Biotechnol. Bioeng. 2016;113: 1244-1250. © 2015 Wiley Periodicals, Inc.

Nutritional interventions to prevent and treat osteoarthritis. Part II: focus on micronutrients and supportive nutraceuticals.

Osteoarthritis (OA) is the most common cause of musculoskeletal disability in the elderly, and it places an enormous economic burden on society, which will remain a major health care challenge with an aging population. Management of OA is primarily focused on palliative relief using agents such as nonsteroidal anti-inflammatory drugs (NSAID) and analgesics. However, such an approach is limited by a narrow therapeutic focus that fails to address the progressive and multimodal nature of OA. Given the favorable safety profile of most nutritional interventions, identifying disease-modifying pharmaconutrients capable of improving symptoms and also preventing, slowing, or even reversing the degenerative process in OA should remain an important paradigm in translational and clinical research. The goals of pharmaconutrition for metabolic optimization are to drive biochemical reactions in a desired direction and to meet health condition-specific metabolic demands. Applying advances in nutritional science to musculoskeletal medicine remains challenging, given the fluid and dynamic nature of the field, along with a rapidly developing regulatory climate over manufacturing and commerce requirements. The purpose of this article is to review the available literature on effectiveness and potential mechanism for OA of micronutrient vitamins; minerals; glycosaminoglycans; avocado-soybean unsaponifiable fractions; methylsulfonylmethane; s-adenosylmethionine; undenatured and hydrolyzed collagen preparations; phytoflavonoid compounds found in fruits, vegetables, spices, teas, and nuts; and other nutrients on the horizon. There also is a discussion on the concept of rational polysupplementation via the strategic integration of multiple nutraceuticals with potential complementary mechanisms for improving outcomes in OA. As applied nutritional science evolves, it will be important to stay on the forefront of proteomics, metabolomics, epigenetics, and nutrigenomics, because they hold enormous potential for developing novel therapeutic and prognostic breakthroughs in many areas of medicine, including OA.

The commercial harvest of devil's claw (Harpagophytum spp.) in southern Africa: the devil's in the details.

Devil's claw is the common name for two species in the genus Harpagophytum. Their root extracts contain the iridoid glycoside, harpagoside, which has been found to be effective in the treatment of degenerative rheumatoid arthritis, osteoarthritis, tendonitis, kidney inflammation, and heart disease. Most of the world's supply comes from Namibia, with lesser amounts from South Africa and Botswana. In 2002, the peak year of export, 1018 tonnes of dried tubers were exported from southern Africa, representing the harvest of millions of plants. In 2001, sales in Germany were estimated at 30 M euros, accounting for 74% of the prescriptions for rheumatism. Harvest has improved income levels in marginalized communities but it has also raised questions of sustainability. In 2000, recommendations were made to the Convention on the International Trade in Endangered Species (CITES) to add devil's claw to Appendix II. In 2004, the proposal was formally withdrawn due to the efforts of the range states to address sustainability issues. Replacing wild collection with cultivation has generated a debate on the positive and negative effects on harvester income and rural farmers. Successful cultivation efforts have involved micropropagation techniques and growing the plant without water or fertilizers. The governments of the main range states are working with local communities to develop policies and regulations to protect the species and to determine a sustainable harvest.