miR146a up-regulation is involved in small HA oligosaccharides-induced pro-inflammatory response in human chondrocytes

BackgroundSmall HA fragments are produced during cartilage degradation and their role seems to be preponderant during pathologies in which cartilage injury contribute to trigger and perpetuate the inflammatory mechanism.Several reports have increasingly shown that MicroRNAs (miRs), a small non-coding mRNAs are involved in the regulation of multiple biological processes, including cell proliferation and inflammation response in different pathologies, among them miR146a seems to be involved in inflammatory processes.MethodsStarting by these evidences we investigated the levels of miR146a and its correlation with inflammatory mediators in an experimental model of 6-mer HA-induced inflammatory response in human cultured chondrocytes.ResultsTreatment of chondrocytes with 6-mer HA showed up-regulation in inflammation parameters such as TLR-4, and CD44 receptors activation, IL-6, IL-1β and MMP-13 mRNA expression and proteins production, as well as NF-kB activation. We also revealed an up-regulation of miR146a. Transfection with a miR146a mimic or miR146a inhibitor produced the following effects: chondrocytes receiving miR146a mimic and then 6-mer HA significantly reduced inflammatory cytokines and MMP-13, while exposition of chondrocytes with miR146a inhibitor and then the 6-mer HA incremented the activity of damaging cytokines and MMP13. Expression of CD44 receptor was not affected by miR-146a treatments, while TLR-4 expression and NF-kB activation were modified.ConclusionsWe concluded that up-regulation of miR146a occurred in 6-mer HA-induced inflammation response may reduce the inflammatory cascade by modulating TLR-4 and NF-kB activation.General significanceThese results could be useful in develop new therapeutic strategies with the aim to reduce OA and RA incidence.     

Distribution and changes of glycoconjugates in rat colonic mucosa during development. A histochemical study using lectins

A study was made of the modifications of glycoconjugates in rat colonic mucosa during development. Sections of the caecum, and proximal and distal portions of the colon from Sprague Dawley rats at different stages of development (embryos, fetuses, suckling, weaning and adult rats) were examined. The sections were incubated with a battery of eight fluoresceinated lectins: DBA, SBA, WGA, LFA, PNA, GS-I, UEA-I and Con A. Some sections were treated with neuraminidase, and others were submitted to sequential saponification-neuraminidase treatment prior to incubation with the lectin (WGA, PNA or LFA). The intensity of the fluorescence was evaluated and graded from absent (-) to very positive (4+). Gradual and progressive changes were seen in colonic glycoconjugates during development. These changes revealed a unique developmental pattern for each lectin, which was independent for each cellular compartment (goblet cells, luminal surface and supranuclear region). Local and regional differences, observed between the different colonic sections, were already present from early stages of development. Moreover, our study showed that for several glycoconjugates, the differentiation process in colonic mucosa began in the distal region and continued through to the proximal region, the former being the first to reach the adult pattern. In the caecum, some lectins maintained a fetal pattern throughout all the periods of development up to the adult stage.     

Homecare monitoring system: A technical proposal for the safety of the elderly experimented in an Alzheimer's care unit

Today, the monitoring of dependent people is a real challenge. This is mainly due to the increase in the number of elderly and reduction of medical staff. However, works done in recent years in France are faced with obstacle of clinical and industrial validation. In this paper, we propose a monitoring system for dependent persons living alone at home or in an institution 24 h/24. This system is based on a presence multisensor network deployed in the living environment of the monitored person coupled with a wireless identification system. A learning algorithm is implemented in order to define a personalized behavioural model (motions deviation, nocturnal activity, falls, mobility). This model allows the nursing staff monitoring the behaviour through a web application accessed remotely, and also intervention in case of dangerous situations thanks to an alert system. In further, we describe the experiments conducted at the Caussade Local Hospital in France with Alzheimer patients and we present the results of data obtained. Finally, we propose the way for an economic model that would allow to industrialize the system with prospects of clinical validation on a larger cohort of patients.     

Assessment of a 1H high-resolution magic angle spinning NMR spectroscopy procedure for free sugars quantification in intact plant tissue

In most plants, sucrose is the primary product of photosynthesis, the transport form of assimilated carbon, and also one of the main factors determining sweetness in fresh fruits. Traditional methods for sugar quantification (mainly sucrose, glucose and fructose) require obtaining crude plant extracts, which sometimes involve substantial sample manipulation, making the process time-consuming and increasing the risk of sample degradation. Here, we describe and validate a fast method to determine sugar content in intact plant tissue by using high-resolution magic angle spinning nuclear magnetic resonance spectroscopy (HR-MAS NMR). The HR-MAS NMR method was used for quantifying sucrose, glucose and fructose in mesocarp tissues from melon fruits (Cucumis melo var. reticulatus and Cucumis melo var. cantalupensis). The resulting sugar content varied among individual melons, ranging from 1.4 to 7.3 g of sucrose, 0.4–2.5 g of glucose; and 0.73–2.83 g of fructose (values per 100 g fw). These values were in agreement with those described in the literature for melon fruit tissue, and no significant differences were found when comparing them with those obtained using the traditional, enzymatic procedure, on melon tissue extracts. The HR-MAS NMR method offers a fast (usually <30 min) and sensitive method for sugar quantification in intact plant tissues, it requires a small amount of tissue (typically 50 mg fw) and avoids the interferences and risks associated with obtaining plant extracts. Furthermore, this method might also allow the quantification of additional metabolites detectable in the plant tissue NMR spectrum.     

A Novel Immunomodulatory Hemocyanin from the Limpet Fissurella latimarginata Promotes Potent Anti-Tumor Activity in Melanoma

Hemocyanins, the huge oxygen-transporting glycoproteins of some mollusks, are used as immunomodulatory proteins with proven anti-cancer properties. The biodiversity of hemocyanins has promoted interest in identifying new anti-cancer candidates with improved immunological properties. Hemocyanins promote Th1 responses without known side effects, which make them ideal for long-term sustained treatment of cancer. In this study, we evaluated a novel hemocyanin from the limpet/gastropod Fissurella latimarginata (FLH). This protein has the typical hollow, cylindrical structure of other known hemocyanins, such as the keyhole limpet hemocyanin (KLH) and the Concholepas hemocyanin (CCH). FLH, like the KLH isoforms, is composed of a single type of polypeptide with exposed N- and O-linked oligosaccharides. However, its immunogenicity was significantly greater than that of KLH and CCH, as FLH induced a stronger humoral immune response and had more potent anti-tumor activity, delaying tumor growth and increasing the survival of mice challenged with B16F10 melanoma cells, in prophylactic and therapeutic settings. Additionally, FLH-treated mice demonstrated increased IFN-γ production and higher numbers of tumor-infiltrating CD4⁺ lymphocytes. Furthermore, in vitro assays demonstrated that FLH, but not CCH or KLH, stimulated the rapid production of pro-inflammatory cytokines (IL-6, IL-12, IL-23 and TNF-α) by dendritic cells, triggering a pro-inflammatory milieu that may explain its enhanced immunological activity. Moreover, this effect was abolished when deglycosylated FLH was used, suggesting that carbohydrates play a crucial role in the innate immune recognition of this protein. Altogether, our data demonstrate that FLH possesses increased anti-tumor activity in part because it activates a more potent innate immune response in comparison to other known hemocyanins. In conclusion, FLH is a potential new marine adjuvant for immunization and possible cancer immunotherapy.