The chrondoprotective actions of a natural product are associated with the activation of IGF-1 production by human chondrocytes despite the presence of IL-1β

BACKGROUND: Cartilage loss is a hallmark of arthritis and follows activation of catabolic processes concomitant with a disruption of anabolic pathways like insulin-like growth factor 1 (IGF-1). We hypothesized that two natural products of South American origin, would limit cartilage degradation by respectively suppressing catabolism and activating local IGF-1 anabolic pathways. One extract, derived from cat's claw (Uncaria guianensis, vincaria(®)), is a well-described inhibitor of NF-κB. The other extract, derived from the vegetable Lepidium meyenii (RNI 249), possessed an uncertain mechanism of action but with defined ethnomedical applications for fertility and vitality. METHODS: Human cartilage samples were procured from surgical specimens with consent, and were evaluated either as explants or as primary chondrocytes prepared after enzymatic digestion of cartilage matrix. Assessments included IGF-1 gene expression, IGF-1 production (ELISA), cartilage matrix degradation and nitric oxide (NO) production, under basal conditions and in the presence of IL-1β. RESULTS: RNI 249 enhanced basal IGF-1 mRNA levels in human chondrocytes by 2.7 fold, an effect that was further enhanced to 3.8 fold by co-administration with vincaria. Enhanced basal IGF-1 production by RNI 249 alone and together with vincaria, was confirmed in both explants and in primary chondrocytes (P <0.05). As expected, IL-1β exposure completely silenced IGF-1 production by chondrocytes. However, in the presence of IL-1β both RNI 249 and vincaria protected IGF-1 production in an additive manner (P <0.01) with the combination restoring chondrocyte IGF-1 production to normal levels. Cartilage NO production was dramatically enhanced by IL-1β. Both vincaria and RNI 249 partially attenuated NO production in an additive manner (p < 0.05). IL-1β – induced degradation of cartilage matrix was quantified as glycosaminoglycan release. Individually RNI 249 or vincaria, prevented this catabolic action of IL-1β. CONCLUSION: The identification of agents that activate the autocrine production of IGF-1 in cartilage, even in the face of suppressive pro-inflammatory, catabolic cytokines like IL-1β, represents a novel therapeutic approach to cartilage biology. Chondroprotection associated with prevention of the catabolic events and the potential for sustained anabolic activity with this natural product suggests that it holds significant promise in the treatment of debilitating joint diseases

A Live-Attenuated HSV-2 ICP0− Virus Elicits 10 to 100 Times Greater Protection against Genital Herpes than a Glycoprotein D Subunit Vaccine

Glycoprotein D (gD-2) is the entry receptor of herpes simplex virus 2 (HSV-2), and is the immunogen in the pharmaceutical industry's lead HSV-2 vaccine candidate. Efforts to prevent genital herpes using gD-2 subunit vaccines have been ongoing for 20 years at a cost in excess of $100 million. To date, gD-2 vaccines have yielded equivocal protection in clinical trials. Therefore, using a small animal model, we sought to determine if a live-attenuated HSV-2 ICP0− virus would elicit better protection against genital herpes than a gD-2 subunit vaccine. Mice immunized with gD-2 and a potent adjuvant (alum+monophosphoryl lipid A) produced high titers of gD-2 antibody. While gD-2-immunized mice possessed significant resistance to HSV-2, only 3 of 45 gD-2-immunized mice survived an overwhelming challenge of the vagina or eyes with wild-type HSV-2 (MS strain). In contrast, 114 of 115 mice immunized with a live HSV-2 ICP0− virus, 0ΔNLS, survived the same HSV-2 MS challenges. Likewise, 0ΔNLS-immunized mice shed an average 125-fold less HSV-2 MS challenge virus per vagina relative to gD-2-immunized mice. In vivo imaging demonstrated that a luciferase-expressing HSV-2 challenge virus failed to establish a detectable infection in 0ΔNLS-immunized mice, whereas the same virus readily infected naïve and gD-2-immunized mice. Collectively, these results suggest that a HSV-2 vaccine might be more likely to prevent genital herpes if it contained a live-attenuated HSV-2 virus rather than a single HSV-2 protein

Precision restoration: a necessary approach to foster forest recovery in the 21st century

We thank S. Tabik, E. Guirado, and Garnata Drone SL for fruitful debates about the application of remote sensing and artificial intelligence in restoration. E. McKeown looked over the English version of the manuscript. Original drawings were made by J. D. Guerrero. This work was supported by projects RESISTE (P18-RT-1927) from the Consejeria de Economia, Conocimiento, y Universidad from the Junta de Andalucia, and AVA201601.19 (NUTERA-DE I), DETECTOR (A-RNM-256-UGR18), and AVA2019.004 (NUTERA-DE II), cofinanced (80%) by the FEDER Program. F.M.-R. acknowledges the support of the Agreement 4580 between OTRI-UGR and the city council of La Zubia. We thank an anonymous reviewer for helpful comments that improved the manuscript.Forest restoration is currently a primary objective in environmental management policies at a global scale, to the extent that impressive initiatives and commitments have been launched to plant billions of trees. However, resources are limited and the success of any restoration effort should be maximized. Thus, restoration programs should seek to guarantee that what is planted today will become an adult tree in the future, a simple fact that, however, usually receives little attention. Here, we advocate for the need to focus restoration efforts on an individual plant level to increase establishment success while reducing negative side effects by using an approach that we term “precision forest restoration” (PFR). The objective of PFR will be to ensure that planted seedlings or sowed seeds will become adult trees with the appropriate landscape configuration to create functional and self-regulating forest ecosystems while reducing the negative impacts of traditional massive reforestation actions. PFR can take advantage of ecological knowledge together with technologies and methodologies from the landscape scale to the individual- plant scale, and from the more traditional, low-tech approaches to the latest high-tech ones. PFR may be more expensive at the level of individual plants, but will be more cost-effective in the long term if it allows for the creation of resilient forests able to providemultiple ecosystemservices. PFR was not feasible a few years ago due to the high cost and low precision of the available technologies, but it is currently an alternative that might reformulate a wide spectrum of ecosystem restoration activities.Junta de Andalucia P18-RT-1927European Commission AVA201601.19 A-RNM-256-UGR18 AVA2019.004OTRI-UGR 4580city council of La Zubia 458

Analysis of protein carbonylation - pitfalls and promise in commonly used methods

Abstract Oxidation of proteins has received a lot of attention in the last decades due to the fact that they have been shown to accumulate and to be implicated in the progression and the patho-physiology of several diseases such as Alzheimer, coronary heart diseases, etc. This has also resulted in the fact that research scientist became more eager to be able to measure accurately the level of oxidized protein in biological materials, and to determine the precise site of the oxidative attack on the protein, in order to get insights into the molecular mechanisms involved in the progression of diseases. Several methods for measuring protein carbonylation have been implemented in different laboratories around the world. However, to date no methods prevail as the most accurate, reliable and robust. The present paper aims at giving an overview of the common methods used to determine protein carbonylation in biological material as well as to highlight the limitations and the potential. The ultimate goal is to give quick tips for a rapid decision making when a method has to be selected and taking into consideration the advantage and drawback of the methods