Den goda lagstiftningen: en kritisk diskursanalys av proposition 1971:51 ny rennäringslag

The exploitation of Sápmi, Sami land, began about 400 years ago. With a post-colonial approach, I, in this essay analyse the discourses behind the law that regulates reindeer herding. With a critical discourse analysis of a government bill (proposition 1971:51) with post-colonial and Marxist theories, I show that discourses of profit and development influence the way Sami culture is perceived. In prop. 1971:51 Sami culture is presumed to be endangered, due to the supposed economic inefficiency of reindeer herding. To save Sami culture, proposition 1971:51 suggests that the reindeer herding should be rationalized. I argue that these discourses are being influenced by a bigger capitalistic discourse of profit

Staten och kapitalet: en kritisk diskursanalys av prop. 1988:92 ny minerallag

One of the biggest threats against reindeer herding and Sami culture is the exploitation of natural resources in Sápmi, Sami land (Samiskt informationscentrum a). Mines require large areas of land, where the reindeer usually feed. With a post-colonial approach, I, in this essay analyse the discourses behind the law that regulates mines and mineral extraction. With a critical discourse analysis of a government bill (proposition 1988:92) with post-colonial and Marxist theories, I show that discourses of profit succeed over discourses representing Sami interests

An Acute Increase of Dietary Protein Intake Elicits Positive Cellular Metabolic Adaptations in Healthy Males

There is emerging literature demonstrating that restricting dietary carbohydrate (CHO) intake might upregulate cellular markers of mitochondrial biogenesis. Mitochondria quantity and density has been linked with increased endurance performance, reduction in type 2 diabetes and improved insulin sensitivity. A number of transcriptional cellular markers have been identified as key regulators of this process. PURPOSE: To determine the influence of 7 days dietary manipulation on resting metabolic rate (RMR), body composition and transcriptional markers of mitochondrial biogenesis. METHOD: Forty-six healthy male participants (mean ± SD; age (years), body mass (kg), height (cm); 28 ± 5, 75.6 ± 11.1, 178.0 ± 4.9, respectively) were recruited and randomised to one of four conditions: energy matched high protein (PRO-EM), energy restricted high protein (PRO-ER), energy matched high carbohydrate (CHO-EM) or energy restricted high carbohydrate (CHO-ER). Macronutrient ratios (PRO:CHO:FAT) of 40:30:30 and 60:10:30 were used for high protein and high carbohydrate conditions, respectively. Calorific intake for energy restricted groups was matched to RMR. Participants visited the laboratory on 3 occasions across 15 days. On days 0, 7 and 15 participants completed assessments of body composition (DEXA) and RMR (indirect calorimetry), prior to providing a muscle biopsy from the vastus lateralis for later analysis of transcriptional markers via real-time polymerase chain reaction. Between days 1 & 7 and 7 & 14 participants consumed their habitual and prescribed diets, respectively. Laboratory testing was completed following an overnight fast and at the same time of day on each occasion. RESULTS: No difference in RMR was observed in any group across all time points. AMPK, PGC-1a, SIRT1 and PPAR expression was increased in the PRO-ER group (1.32, 1.20, 1.45 and 1.41 fold, respectively). Transcriptional markers were not affected in either CHO group. The CHO-ER group demonstrated a greater loss in lean mass relative to the PRO-EM (-2.22 vs -0.35%,) and body mass loss relative to both CHO-EM and PRO-EM (-2.85 vs -0.95 vs -1.47%) (P < 0.05). CONCLUSION: A restriction energy intake combined with increased protein consumption for 7 days increases transcriptional markers of mitochondrial biogenesis

An unexpected link between fatty acid synthase and cholesterol synthesis in proinflammatory macrophage activation

Different immune activation states require distinct metabolic features and activities in immune cells. For instance, inhibition of fatty acid synthase (FASN), which catalyzes the synthesis of long-chain fatty acids, prevents the proinflammatory response in macrophages; however, the precise role of this enzyme in this response remains poorly defined. Consistent with previous studies, we found here that FASN is essential for lipopolysaccharide-induced, Toll-like receptor (TLR)-mediated macrophage activation. Interestingly, only agents that block FASN upstream of acetoacetyl-CoA synthesis, including the well-characterized FASN inhibitor C75, inhibited TLR4 signaling, while those acting downstream had no effect. We found that acetoacetyl-CoA could overcome C75's inhibitory effect, whereas other FASN metabolites, including palmitate, did not prevent C75-mediated inhibition. This suggested an unexpected role for acetoacetyl-CoA in inflammation that is independent of its role in palmitate synthesis. Our evidence further suggested that acetoacetyl-CoA arising from FASN activity promotes cholesterol production, indicating a surprising link between fatty acid synthesis and cholesterol synthesis. We further demonstrate that this process is required for TLR4 to enter lipid rafts and facilitate TLR4 signaling. In conclusion, we have uncovered an unexpected link between FASN and cholesterol synthesis that appears to be required for TLR signal transduction and proinflammatory macrophage activation

Pharmacological validation of targets regulating CD14 during macrophage differentiation

The signalling receptor for LPS, CD14, is a key marker of, and facilitator for, pro-inflammatory macrophage function. Pro-inflammatory macrophage differentiation remains a process facilitating a broad array of disease pathologies, and has recently emerged as a potential target against cytokine storm in COVID19. Here, we perform a whole-genome CRISPR screen to identify essential nodes regulating CD14 expression in myeloid cells, using the differentiation of THP-1 cells as a starting point. This strategy uncovers many known pathways required for CD14 expression and regulating macrophage differentiation while additionally providing a list of novel targets either promoting or limiting this process. To speed translation of these results, we have then taken the approach of independently validating hits from the screen using well-curated small molecules. In this manner, we identify pharmacologically tractable hits that can either increase CD14 expression on non-differentiated monocytes or prevent CD14 upregulation during macrophage differentiation. An inhibitor for one of these targets, MAP2K3, translates through to studies on primary human monocytes, where it prevents upregulation of CD14 following M-CSF induced differentiation, and pro-inflammatory cytokine production in response to LPS. Therefore, this screening cascade has rapidly identified pharmacologically tractable nodes regulating a critical disease-relevant process

Human rhinovirus promotes STING trafficking to replication organelles to promote viral replication

Human rhinovirus (HRV), like coronavirus (HCoV), are positive-strand RNA viruses that cause both upper and lower respiratory tract illness, with their replication facilitated by concentrating RNA-synthesizing machinery in intracellular compartments made of modified host membranes, referred to as replication organelles (ROs). Here we report a non-canonical, essential function for stimulator of interferon genes (STING) during HRV infections. While the canonical function of STING is to detect cytosolic DNA and activate inflammatory responses, HRV infection triggers the release of STIM1-bound STING in the ER by lowering Ca2+, thereby allowing STING to interact with phosphatidylinositol 4-phosphate (PI4P) and traffic to ROs to facilitates viral replication and transmission via autophagy. Our results thus hint a critical function of STING in HRV viral replication and transmission, with possible implications for other RO-mediated RNA viruses

Differential recognition of HIV-stimulated IL-1? and IL-18 secretion through NLR and NAIP signalling in monocyte-derived macrophages

Macrophages are important drivers of pathogenesis and progression to AIDS in HIV infection. The virus in the later phases of the infection is often predominantly macrophage-tropic and this tropism contributes to a chronic inflammatory and immune activation state that is observed in HIV patients. Pattern recognition receptors of the innate immune system are the key molecules that recognise HIV and mount the inflammatory responses in macrophages. The innate immune response against HIV-1 is potent and elicits caspase-1-dependent pro-inflammatory cytokine production of IL-1β and IL-18. Although, NLRP3 has been reported as an inflammasome sensor dictating this response little is known about the pattern recognition receptors that trigger the “priming” signal for inflammasome activation, the NLRs involved or the HIV components that trigger the response. Using a combination of siRNA knockdowns in monocyte derived macrophages (MDMs) of different TLRs and NLRs as well as chemical inhibition, it was demonstrated that HIV Vpu could trigger inflammasome activation via TLR4/NLRP3 leading to IL-1β/IL-18 secretion. The priming signal is triggered via TLR4, whereas the activation signal is triggered by direct effects on Kv1.3 channels, causing K+ efflux. In contrast, HIV gp41 could trigger IL-18 production via NAIP/NLRC4, independently of priming, as a one-step inflammasome activation. NAIP binds directly to the cytoplasmic tail of HIV envelope protein gp41 and represents the first non-bacterial ligand for the NAIP/NLRC4 inflammasome. These divergent pathways represent novel targets to resolve specific inflammatory pathologies associated with HIV-1 infection in macrophages

Improving the efficiency and effectiveness of an industrial SARS-CoV-2 diagnostic facility.

On 11th March 2020, the UK government announced plans for the scaling of COVID-19 testing, and on 27th March 2020 it was announced that a new alliance of private sector and academic collaborative laboratories were being created to generate the testing capacity required. The Cambridge COVID-19 Testing Centre (CCTC) was established during April 2020 through collaboration between AstraZeneca, GlaxoSmithKline, and the University of Cambridge, with Charles River Laboratories joining the collaboration at the end of July 2020. The CCTC lab operation focussed on the optimised use of automation, introduction of novel technologies and process modelling to enable a testing capacity of 22,000 tests per day. Here we describe the optimisation of the laboratory process through the continued exploitation of internal performance metrics, while introducing new technologies including the Heat Inactivation of clinical samples upon receipt into the laboratory and a Direct to PCR protocol that removed the requirement for the RNA extraction step. We anticipate that these methods will have value in driving continued efficiency and effectiveness within all large scale viral diagnostic testing laboratories

Pharmacological validation of targets regulating CD14 during macrophage differentiation

The signalling receptor for LPS, CD14, is a key marker of, and facilitator for, pro-inflammatory macrophage function. Pro-inflammatory macrophage differentiation remains a process facilitating a broad array of disease pathologies, and has recently emerged as a potential target against cytokine storm in COVID19. Here, we perform a whole-genome CRISPR screen to identify essential nodes regulating CD14 expression in myeloid cells, using the differentiation of THP-1 cells as a starting point. This strategy uncovers many known pathways required for CD14 expression and regulating macrophage differentiation while additionally providing a list of novel targets either promoting or limiting this process. To speed translation of these results, we have then taken the approach of independently validating hits from the screen using well-curated small molecules. In this manner, we identify pharmacologically tractable hits that can either increase CD14 expression on non-differentiated monocytes or prevent CD14 upregulation during macrophage differentiation. An inhibitor for one of these targets, MAP2K3, translates through to studies on primary human monocytes, where it prevents upregulation of CD14 following M-CSF induced differentiation, and pro-inflammatory cytokine production in response to LPS. Therefore, this screening cascade has rapidly identified pharmacologically tractable nodes regulating a critical disease-relevant process