Neuroinflammatory response to lipopolysaccharide is exacerbated in mice genetically deficient in cyclooxygenase-2

<p>Abstract</p> <p>Background</p> <p>Cyclooxygenases (COX) -1 and -2 are key mediators of the inflammatory response in the central nervous system. Since COX-2 is inducible by inflammatory stimuli, it has been traditionally considered as the most appropriate target for anti-inflammatory drugs. However, the specific roles of COX-1 and COX-2 in modulating a neuroinflammatory response are unclear. Recently, we demonstrated that COX-1 deficient mice show decreased neuroinflammatory response and neuronal damage in response to lipopolysaccharide (LPS).</p> <p>Methods</p> <p>In this study, we investigated the role of COX-2 in the neuroinflammatory response to intracerebroventricular-injected LPS (5 μg), a model of direct activation of innate immunity, using COX-2 deficient (COX-2^-/-) and wild type (COX-2^+/+) mice, as well as COX-2^+/+mice pretreated for 6 weeks with celecoxib, a COX-2 selective inhibitor.</p> <p>Results</p> <p>Twenty-four hours after LPS injection, COX-2^-/-mice showed increased neuronal damage, glial cell activation, mRNA and protein expression of markers of inflammation and oxidative stress, such as cytokines, chemokines, iNOS and NADPH oxidase. Brain protein levels of IL-1β, NADPH oxidase subunit p67^phox, and phosphorylated-signal transducer and activator of transcription 3 (STAT3) were higher in COX-2^-/-and in celecoxib-treated mice, compared to COX-2^+/+mice. The increased neuroinflammatory response in COX-2^-/-mice was likely mediated by the upregulation of STAT3 and suppressor of cytokine signaling 3 (SOCS3).</p> <p>Conclusion</p> <p>These results show that inhibiting COX-2 activity can exacerbate the inflammatory response to LPS, possibly by increasing glial cells activation and upregulating the STAT3 and SOCS3 pathways in the brain.</p

Stakeholder Participation in Planning of a Sustainable and Competitive Tourism Destination: The Genoa Integrated Action Plan

The outbreak of COVID-19 confronted the international community with critical health, social, and economic challenges. Travel and tourism were among the hardest affected sectors. In 2020 and 2021 new travel trends emerged, emphasizing local destinations, short distances, and consequently, lower-carbon transportation (proximity tourism). Post-pandemic recovery represents an opportunity to bounce back better by rethinking the sector’s economic model for the sake of sustainability and innovation. This paper disseminates the research that led to the structuring of guidelines for a breakthrough and inclusive municipal-level action plan for the promotion of sustainable tourism, as part of the Tourism Friendly Cities project. An operational methodology is discussed here, whereby key stakeholder participation, conceptualized through a sextuple helix model, is the foundation of the planning process. A small-scale action and a qualitative assessment tool of the participatory process are also illustrated. The proposed methodology corroborates the vast positive effects deriving from stakeholder participation in terms of trust, ownership, planning quality, innovativeness and sustainability of interventions. In applying the methodology, although the digital framework was evaluated positively in terms of the number of participants that could be involved, data collection, and confidentiality of activities, the evaluation shows that hybrid modes of participation are more desirable

The role of non-hematopoietic stromal cells in the persistence of inflammation

Inflammation results from the complex interaction between hematopoietic and stromal cells and growing evidence supports a key role for the stroma in driving the switch from acute resolving to persistence in chronic inflammatory diseases. Stromal cells have also been shown to play a critical role in cancer biology, being involved in cancer growth, dissemination, and inhibition of the autologous immune response, ultimately favoring persistence and metastatic spread. Similarly, blood and lymphatic endothelial cells contribute to tissue homeostasis during physiological inflammation but also lead to discorded leukocyte and tumor cell accumulation in pathological inflammation and cancer. This review aims to summarize the role that pathogenic stroma plays in the pathogenesis of diseases such as cancer and chronic inflammation

Different Serum Free Fatty Acid Profiles in NAFLD Subjects and Healthy Controls after Oral Fat Load

Background: Free fatty acid (FFA) metabolism can impact on metabolic conditions, such as obesity and nonalcoholic fatty liver disease (NAFLD). This work studied the increase in total FFA shown in NAFLD subjects to possibly characterize which fatty acids significantly accounted for the whole increase. Methods: 21 patients with NAFLD were selected according to specified criteria. The control group consisted of nine healthy subjects. All subjects underwent an oral standard fat load. Triglycerides; cholesterol; FFA; glucose and insulin were measured every 2 h with the determination of fatty acid composition of FFA. Results: higher serum FFA levels in NAFLD subjects are mainly due to levels of oleic, palmitic and linoleic acids at different times. Significant increases were shown for docosahexaenoic acid, linolenic acid, eicosatrienoic acid, and arachidonic acid, although this was just on one occasion. In the postprandial phase, homeostatic model assessment HOMA index positively correlated with the ω3/ω6 ratio in NAFLD patients. Conclusions: the higher serum levels of FFA in NAFLD subjects are mainly due to levels of oleic and palmitic acids which are the most abundant circulating free fatty acids. This is almost exactly corresponded with significant increases in linoleic acid. An imbalance in the n-3/n-6 fatty acids ratio could modulate postprandial responses with more pronounced effects in insulin-resistant subjects, such as NAFLD patients

A functional genetic toolbox for human tissue-derived organoids.

Funder: Alzheimers Research UK Stem Cell Research CentreHuman organoid systems recapitulate key features of organs offering platforms for modelling developmental biology and disease. Tissue-derived organoids have been widely used to study the impact of extrinsic niche factors on stem cells. However, they are rarely used to study endogenous gene function due to the lack of efficient gene manipulation tools. Previously, we established a human foetal lung organoid system (Nikolić et al., 2017). Here, using this organoid system as an example we have systematically developed and optimised a complete genetic toolbox for use in tissue-derived organoids. This includes 'Organoid Easytag' our efficient workflow for targeting all types of gene loci through CRISPR-mediated homologous recombination followed by flow cytometry for enriching correctly-targeted cells. Our toolbox also incorporates conditional gene knock-down or overexpression using tightly-inducible CRISPR interference and CRISPR activation which is the first efficient application of these techniques to tissue-derived organoids. These tools will facilitate gene perturbation studies in tissue-derived organoids facilitating human disease modelling and providing a functional counterpart to many on-going descriptive studies, such as the Human Cell Atlas Project