Ex vivo culture of keratinocytes on papillary and reticular dermal layers remodels skin explants differently: towards improved wound care

In this study, we characterised the efect that seeding keratinocytes on the papillary and reticular dermis had on the extracellular matrix and tissue integrity ex vivo. Human skin explants from consented patients (n=6) undergoing routine surgery were cultured at a liquid–air interface, dermal-side up, and autologous keratinocytes seeded on the exposed papillary or reticular layer. After 7–21 days, histological and immunohistochemical evaluation of the morphology and extracellular matrix was performed. While the dermis remained robust in all explants cultures, keratinocytes seeded on the papillary layer showed less tissue infltration and remodelling and formed clusters across the tissue. In contrast, keratinocytes seeded on the reticular layer infltrated the tissue homogenously with an intact single-cell-layer surface coverage and structural changes characterised by increased deposition of ground substance, glycosaminoglycans, and collagen VII in 14 days. In addition, while the papillary section showed more new laminin deposition by 14 days than the reticular section, the latter expressed more connexin 43. These diferences in re-epithelialisation and extracellular matrix characteristics suggest that wound depth and graft thickness may play a key role in wound healing and indicate that ECM characteristics should be factored in when designing biomaterials for wound applications and in the selection of recipient sites when using cells for grafting

Artificial Intelligence approaches for predicting Harmful Algal Blooms (HABs)

This is the final version.Final Report to Department for Business, Energy and Industrial StrategyHABs can produce toxins, which accumulate in filter-feeding shellfish and intoxicate human consumers. The toxins are heat stable and so can't be destroyed by freezing and/or cooking. Under current regulations shellfish toxin monitoring is effectively retrospective: regulators sample, await results, and if the regulatory threshold is breached there is an investigation into the amount of shellfish harvested since the sample was taken, which might then result in a full-scale food chain product recall. By gathering high resolution field monitoring data using novel qPCR and lateral-flow (LF) techniques, we planned to refine and validate a computer model for predicting HABs caused by Dinophysis species. One of the aims of the project was to use the higher resolution data collected as part of the project to train the model towards a more accurate forecast in respect of breaches in the Dinophysis toxin threshold up to 6-8 weeks ahead. The model would then aid planning decisions for harvesting and will save costly recalls and protect human health (in this case from Diarrhetic Shellfish Poisoning - DSP). Other strands of the project consisted of use of a Novel monitoring tools, a qPCR for quantifying HAB cell abundance in seawater, and a Lateral Flow testing for quantifying Dinophysis toxins in shellfish, directly in the field. Field data from these novel methods will be validated by an accredited light microscopy technique which enables the cell densities to be quantified in water and by liquid chromatography with tandem mass spectrometry (LC-MS/MS) for validating the shellfish flesh test results from the field.Department for Environment, Food and Rural Affairs (DEFRA

Engaging with History after Macpherson

The Race Relations Amendment Act (2000) identifies a key role for education, and more specifically history, in promoting ‘race equality’ in Britain. In this article Ian Grosvenor and Kevin Myers consider the extent of young people’s current engagement with the history of ‘diversity, change and immigration’ which underpins the commitment to ‘race equality’. Finding that in many of Britain’s schools and universities a singular and exclusionary version of history continues to dominate the curriculum, they go on to consider the reasons for the neglect of multiculturalism. The authors identify the development of an aggressive national identity that depends on the past for its legitimacy and argue that this sense of the past is an important obstacle to future progress

Pharmacological inhibition of Akt and downstream pathways modulates the expression of COX-2 and mPGES-1 in activated microglia

<p>Abstract</p> <p>Background</p> <p>Microglia are considered a major target for modulating neuroinflammatory and neurodegenerative disease processes. Upon activation, microglia secrete inflammatory mediators that contribute to the resolution or to further enhancement of damage in the central nervous system (CNS). Therefore, it is important to study the intracellular pathways that are involved in the expression of the inflammatory mediators. Particularly, the role of the phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) and glycogen synthase kinase-3 (GSK-3) pathways in activated microglia is unclear. Thus, in the present study we investigated the role of Akt and its downstream pathways, GSK-3 and mTOR, in lipopolysaccharide (LPS)-activated primary rat microglia by pharmacological inhibition of these pathways in regard to the expression of cyclooxygenase (COX)-2 and microsomal prostaglandin E synthase-1 (mPGES-1) and to the production of prostaglandin (PG) E₂and PGD₂.</p> <p>Findings</p> <p>We show that inhibition of Akt by the Akt inhibitor X enhanced the production of PGE₂and PGD₂without affecting the expression of COX-2, mPGES-1, mPGES-2 and cytosolic prostaglandin E synthase (cPGES). Moreover, inhibition of GSK-3 reduced the expression of both COX-2 and mPGES-1. In contrast, the mTOR inhibitor rapamycin enhanced both COX-2 and mPGES-1 immunoreactivity and the release of PGE₂and PGD₂. Interestingly, NVP-BEZ235, a dual PI3K/mTOR inhibitor, enhanced COX-2 and reduced mPGES-1 immunoreactivity, albeit PGE₂and PGD₂levels were enhanced in LPS-stimulated microglia. However, this compound also increased PGE₂in non-stimulated microglia.</p> <p>Conclusion</p> <p>Taken together, we demonstrate that blockade of mTOR and/or PI3K/Akt enhances prostanoid production and that PI3K/Akt, GSK-3 and mTOR differently regulate the expression of mPGES-1 and COX-2 in activated primary microglia. Therefore, these pathways are potential targets for the development of novel strategies to modulate neuroinflammation.</p

Short- and mid-term effects on performance, health and qualitative behavioural assessment of Romane lambs in different milk feeding conditions

The common practice of artificially rearing lambs from prolific meat breeds of sheep constitutes a welfare issue due to increased mortality rates and negative health issues. In this multidisciplinary study, we investigated the possible short- and mid-term advantages of artificially feeding fresh ewe's milk instead of commercial milk replacer on lambs' growth, health and welfare. Romane lambs were either separated from their mothers on D3 and fed with Lacaune ewes' milk (LAC, n?=?13) or milk replacer (REP, n?=?15), or they were reared by their mothers (MOT, n =?15). On D45, they were weaned, gathered in single-sex groups until the end of the study on D150. Lamb performance and biomarkers of overall health were assessed by measuring: growth, dirtiness of the perianal area, enteric pathogens in the faeces, total antioxidant status and redox status assessed by plasma reduced glutathione/oxidised glutathione ratio, and immune response after vaccination against chlamydiosis. As an exploratory approach, blood cell transcriptomic profiles were also investigated. Last, qualitative behaviour assessment (QBA) was performed as an integrated welfare criterion. Lacaune ewes' milk and REP never differed in their average daily gain but grew less than MOT lambs in the early suckling period and just after weaning. No effect was detected afterwards. On D30, LAC and REP lambs had lower total antioxidant and higher redox status than MOT lambs but did not differ among themselves. Lacaune ewes' milk and MOT had a cleaner perianal area than REP lambs on D21, while faecal pathogen infection did not vary between the treatment groups. After vaccination, LAC also had a stronger immune response on D90 compared to REP lambs. Transcriptome analysis performed on D150 showed differential gene expression, mainly in relation to inflammatory, immune and cell cycle response, between male lambs of the LAC group and those of the MOT and REP groups. Based on QBA, LAC lambs never differed from MOT lambs in their general activity and varied from REP only on D21; REP lambs were always more agitated than MOT lambs. In conclusion, artificial milk feeding impaired early growth rate, health and emotional state mainly during the milk feeding period and at weaning. Feeding artificially reared lambs with fresh ewe's milk partly mitigated some of the negative effects induced by milk replacer but without achieving the full benefit of being reared by the mother

Using the suture/adhesive strips combination technique for skin closure in an individual with Ehlers–Danlos Syndrome

Combining sutures with adhesive strips to avoid the ‘cheese-wiring’ effect in individuals with fragile skin is a method that has been described previously. Here we demonstrate its application in an individual with Ehlers-Danlos Syndrome. Keywords: Cheese-wire effect, Sutures, Ehlers-Danlos Sydrome, Steri-Strip

Forklift-related lower limb injuries: a retrospective case series study with patient-reported outcome measures (PROMs).

AIM Forklift trucks can cause serious lower limb trauma with long-lasting sequelae to patients. The aim of this study was to analyse a case series of patients with forklift-related injuries over 7 years at a level 1 major trauma centre in the UK and present their patient-reported outcome measures (PROMs) with long-term follow-up. To the best of the authors' knowledge, this is the largest case series study in the UK describing forklift injuries. METHODS Retrospective case note analysis of 19 patients over 7 years. Data including demographics, injury mechanism, pattern of injury, management, length of hospital stay, number of operations and complications were extracted from the notes. We used 'Enneking score' as a validated tool for PROMs. RESULTS Seventeen men and two women with mean age of 47 years; 20% had bilateral injuries and 34% had multi-level fractures. The mean number of theatre sessions was 5.21, while the mean length of hospital stay was 30.10 days. There was one mortality. Twelve patients (63%) required reconstruction with free tissue transfer, with one flap failure. The mean long-term Enneking percentage score was 57.33%. The mean Enneking score for patients in this study is lower than our institute's score for Gustilo 3B, highlighting the gravity of these injuries. CONCLUSION Forklifts can cause grave injuries with massive energy transfer. This study highlights the seriousness of those injuries, thus guiding patient counselling and optimising planning of management