bluuebunny/crossref_metadata_2025_split huggingface dataset:
O15 Transcriptomic profiling reveals regulators of dermal fibroblast state switching
Authors: Kirk Thomas , Ahmed Abubkr , Bensa Tjaša , Tekkala Stavroula , Khan Mohammad , Connelly John , Rognoni Emanuel 
Published: May 2024

Abstract

Introduction and aims
Fibroblasts, the major cell population in connective tissue, are the guardians of the extracellular matrix homeostasis. In adult tissue they exist in a quiescent state that can persist for years, but upon tissue injury, they become rapidly activated and re-enter the cell cycle, followed by deposition and reorganization of the dermal matrix. The fundamental mechanism by which fibroblasts can maintain quiescence and be primed for prompt activation is currently unknown. Deregulations of this process in the skin and other organs is associated with several pathologies including fibrosis or chronic wounds. Notably, rapid fibroblast state switching is also observed during skin development, where a tightly controlled change from a highly proliferative state to a quiescent state promotes extracellular matrix deposition and maturation of the dermal architecture. Understanding the shared mechanisms and differences between development and wound healing will help in the development of tissue regenerative therapies.

Methods
Using dermal development as a model for fibroblast state switching, we integrated several single-cell RNA-Seq and assay for transposase-accessible chromatin-Seq datasets of whole skin during embryonic and postnatal growth. Our novel integrated multiomics single-cell dataset can recapitulate skin developmental changes observed in vivo and reveal the major regulatory networks.

Results
We identified the activating protein (AP)-1 member cJun as the key regulator of fibroblast state switching in skin development and wound repair. Mechanistically, in quiescent adult fibroblasts cJun is highly transcribed, but the protein is constantly ubiquitinated and degraded, priming fibroblasts for rapid activation when needed. Upon injury cJun ubiquitination is reduced, allowing an accumulation of cJun leading to a change in AP-1 dimer composition at the chromatin level, inhibiting fibroblast quiescence and promoting proliferation and activation.

Conclusions
In normal wounds cJun accumulation is inhibited during the advanced (remodelling) wounding phase; however, chronic wounds and fibrotic disease conditions show a continuous and pathological cJun stabilization, which can be targeted through pharmacological intervention.
O20 Dissecting junctional epidermolysis bullosa heterogeneity
Authors: Garcha Priya , Jones Eleri , Ahmed Abubkr Abdelaal , Aumailley Monique , Rognoni Emanuel , Caley Matthew 
Published: May 2024

Abstract

Introduction and aims
Epidermolysis bullosa (EB) is a family of rare, incurable, inherited disorders that are characterized by extreme skin and mucosal fragility. One of the most severe forms is junctional EB (JEB). JEB is caused by mutations in genes encoding the skin basement membrane proteins laminin 332 (α3, β3, γ2), type XVII collagen or integrin α6β4. Despite similarities between the disease pathology of JEB and other forms of EB there is a dramatic reduction in life expectancy for patients with JEB. Patients with JEB rarely survive beyond the first year of life. To investigate this disparity, we have optimized a previously published conditional tamoxifen induced knockout JEB mouse model (Lama3flx/flxK14CreERT). This mouse model recapitulates all key features of JEB, such as thickening of the epidermis and visible blistering owing to the loss of laminin α3 at the basement membrane.

Methods
Single-cell (sc)RNA-Seq analysis was conducted on pooled skin samples from a healthy (control) and JEB mouse model (n = 3 per genotype). Immunofluorescent staining was conducted to confirm scRNA-Seq analysis including markers for skin differentiation (K14, transglutaminase 1, loricrin), proliferation (Ki67), basement membrane (laminin α3, laminin β3, laminin γ2, collagen VII) and immune markers (F4/80, Ly6-G/C, CD3, CD79).

Results
We can see large differences in cellular populations across the two conditions. With increased T- and B-immune-cell subpopulations in the JEB model compared with the healthy control. Utilizing CellChatDB we identified disease-specific signalling pathways that identify activated T- and B-cell populations interacting with JEB macrophage populations via proinflammatory pathways.

Conclusions
We have utilized an in vivo mouse model to gain insight into the molecular and cellular differences in a JEB diseased phenotype. Identifying changes in cellular pathways could be used as a targeted approach to identify a beneficial therapeutic treatment for JEB.
P19 Defining the mechanism of skin barrier restoration by Lactobacillus rhamnosus
Authors: Coppock Lisa , O’Shaughnessy Ryan , O’Neill Catherine 
Published: May 2024

Abstract

Introduction and aims
We aim to investigate the lysate of Lactobacillus rhamnosus GG (LGG), a bacterium previously established as a gut probiotic, and to examine its postulated ability to enhance and restore human skin barrier function. Compromised skin barriers are implicated in various skin conditions including atopic dermatitis. Furthermore, skin barrier function decreases with age leading to dryness and sensitivity. Previous research has shown promising effects of LGG lysates on keratinocytes, including increasing transepidermal electrical resistance, increasing tight junction protein expression, cell migration promotion and mitigation of Staphylococcus aureus toxicity. This study aims to take a closer look at the molecular mechanisms behind the effect of LGG lysates on the skin, including terminal differentiation, lamin dispersal and nuclear degradation in keratinocytes.

Methods
Normal human epithelial keratinocytes and rat epithelial keratinocytes were grown and treated with the LGG lysate. Western blot analysis was used to determine the relative concentrations of tight junction proteins and other differentiation markers. A phosphoproteomic study was performed to identify key pathways activated by the LGG lysate. A Staphylococcus epidermidis lysate control was used to filter out any nonspecific bacterial effects. Immunostaining of nuclear proteins was used to visualize the effects of LGG lysate on lamin dispersal and nuclear degradation. short hairpin RNA filaggrin knockdown normal human keratinocytes were developed to create a ‘barrier impaired’ model to assess ability of the lysate to ‘rescue’ the skin barrier.

Results
The LGG lysate was shown to increase tight junction protein expression in normal human keratinocytes and increase keratinocyte terminal differentiation markers. We present Results that show decreased tight junction protein expression in filaggrin knockdown cells.

Conclusions
These Results show the potential for LGG lysate to enhance and restore skin barrier function and begin to suggest potential mechanisms for this effect.
P20 Characterizing monoamine signalling in human dermal fibroblasts and keratinocytes
Authors: Andriuskevicius Tomas , Parke Brenna , Rhodes Anna , Knight Leigh , Hashemi Parastoo , Higgins Claire A 
Published: May 2024

Abstract

Introduction and aims
Our mental health depends upon a fine balance of monoamines in the brain. Interestingly, there is ample evidence indicating that mental health disorders such as stress or anxiety have an impact on skin condition and appearance. For example, stress induces neurogenic inflammation, which in turn leads to mast cell degranulation in the skin and increases local histamine levels. This increased histamine within the skin exacerbates skin conditions such as eczema and psoriasis, while impacting the skin barrier. In this study, we first sought to investigate monoamine signalling in both skin keratinocytes and fibroblasts, with a specific focus on histamine and serotonin modulation. Next, we investigated mechanisms of release via pharmacological intervention to block corresponding receptors, informed by using single-cell RNA sequencing from existing datasets.

Methods
We adapted a powerful electrochemical technique, fast-scan cyclic voltammetry (FSCV) to codetect monoamine signalling in real-time in vitro. Employing the advanced capabilities of our artificial neural network models, we analysed extensive FSCV colour plot data in real-time to establish neurotransmitter release patterns and understand their characteristics.

Results
Our Results show that both keratinocytes and fibroblasts are capable of releasing serotonin and histamine without external intervention. After administering diphenhydramine, an H1 receptor inhibitor, to the cells, we observed a rise in the frequency and amplitude of both serotonin and histamine signalling compared with the cells that received no treatment. However, bafilomycin, a vacuolar H+ ATPase inhibitor, reduced the signalling frequency significantly. This suggests a potential autoregulation mechanism and interdependency between the two monoamines.

Conclusions
Overall, using analytical Methods, our work shows that skin-resident fibroblasts and keratinocytes can release monoamines including serotonin and histamine. This finding provides an opportunity to examine interventions that facilitate serotonin release, to decrease histamine levels, and improve the skin function and general mental wellbeing.
P21 Advancing dermatological care: a comprehensive evaluation of the steroid-free GL-1 formulation
Authors: Sharma Pragya 
Published: May 2024

Abstract

Introduction and aims
The prolonged use of topical steroids introduces notable dermatological risks, including the onset of permanent stretch marks, bruising, discoloration and vascular abnormalities. This study presents an innovative strategy by formulating GL-1, a steroid-free treatment amalgamating seaweed bioactives, sesame oil and naturally occurring salts and minerals. The objective of this study was to address the limitations of conventional topical steroids and introduce pioneering solutions to dermatological care.

Methods
To assess the safety and efficacy of GL-1 (product 1), a comprehensive battery of in vitro tests was employed. The skin corrosion test (OECD 431) established the noncorrosive nature of GL-1, validating its safety on reconstructed human epidermis. The skin irritation test (OECD 439) demonstrated the nonirritating attributes of GL-1, aligning with the United Nations Globally Harmonized System of Classification and Labelling guidelines. Additionally, the bovine corneal opacity and permeability test (OECD 437) affirmed the noninducing nature of GL-1 in relation to eye irritation or serious eye damage.

Results
Preliminary outcomes from dermal clinical trials indicate promising Results for GL-1. In vitro experiments on HaCaT and human dermal fibroblasts, adult (HDFa) cell lines highlighted the noncytotoxicity of GL-1, contrasting with the ‘Cortopin’, which exhibited cytotoxicity. GL-1 exhibited a proliferative effect in HDFa cells, signifying its potential for regenerative applications. Furthermore, both GL-1 and ‘Cortopin’ displayed antioxidant activity in HaCaT cells, underscoring the additional merits of our formulation.

Conclusions
Our steroid-free formulation, GL-1, demonstrates exceptional safety and efficacy through a meticulous series of in vitro assessments and initial clinical trials. These findings position GL-1 as a promising candidate for diverse dermal applications, offering a secure alternative to traditional topical steroids. The potential impact of GL-1 transcends its therapeutic efficacy, emphasizing its robust safety profile validated through multiple in vitro evaluations.
P25 Identification of early changes in extracellular microRNA biomarkers of scleroderma for point-of-care diagnosis
Authors: Teenan Oliver , Kenaan Ahmad , Delgaldo Francesco , Malaktou Christina , Strid Jessica , Daniels Connor , Ladame Sylvain , Higgins Claire 
Published: May 2024

Abstract

Introduction and aims
Scleroderma is a rare autoimmune disease, characterized by excessive extracellular matrix deposition leading to chronic inflammation and fibrosis. As a progressive disease initially affecting the skin, it presents similarly to Raynaud phenomenon, which delays diagnosis and treatment. Clinical diagnostic tests lack sensitivity, therefore novel Methods of detection and sensitive biomarkers are required to improve diagnosis rates. MicroRNAs are short, stable, noncoding RNAs, often present in the extracellular space, and can be sampled from interstitial skin fluid (ISF) through the employment of hydrogel microneedles. We aim to use this technique to improve upon current clinical gold standards of diagnosing scleroderma, at earlier stages of the disease.

Methods
In this study, we performed next-generation small-RNA sequencing on ISF from healthy individuals, patients prior to a clinical diagnosis of scleroderma [termed very early detection of systemic sclerosis (VEDOSS)], and patients with scleroderma classified as low and high skin score, identifying a panel of microRNA biomarkers for scleroderma diagnosis. Swellable hydrogel microneedles were manufactured in house, applied to excised human tissue and preclinical mouse models to demonstrate the ability to access and quantify extracellular microRNAs in ISF.

Results
Patients who received a VEDOSS diagnosis have increased ISF abundance of miR-379-5p and miR-125a-5p and decreased miR-30e-3p compared with healthy patients. miR-379-5p, miR-363-3p and miR-221-3p are sensitive to the progression of scleroderma and could be used to track the disease with improved sensitivity to autoantibodies in blood or where clinical practices usually require a skin biopsy. Using preclinical models of skin disease, we could differentiate between conditions, with higher sensitivity than blood sampling using our microneedle technology.

Conclusions
By identifying extracellular mircoRNAs, we have shown the potential use of these sensitive biomarkers in a liquid biopsy that can be applied at point-of-care. Peptide nucleic acids complementary to our microRNA biomarkers, will be used to provide real-time on-the-chip readouts of these ­biomarkers.
P26 The effect of interleukin-23 stimulation on gene expression in CD4+ Jurkat cells
Authors: Shellard Elan 
Published: May 2024

Abstract

Introduction and aims
There is now robust evidence that genetic changes along the interleukin (IL)-23 axis increase the risk of developing psoriasis, with CD4+ T cells likely to be pivotal in the disease pathway. Here, we stimulated Jurkat CD4+ T-cell lines that expressed either the risk or protective version of IL23R, identified through genetic studies in psoriasis, and measured the effect in whole-genome expression at different timepoints poststimulation, with the aim of characterizing the downstream effect on the IL-23 pathway.

Methods
The transformed Jurkat CD4+IL12Rβ1+ IL23Rwt/IL23Rmut-R381Q T cells were stimulated with IL-23 protein (10 ng mL−1) for 0, 2, 4, 16 and 24 h, with triplicates used for each timepoint. RNA sequencing was performed by Novogene, and statistical analyses was performed using negative binomial generalized linear models and then a likelihood-ratio test to remove condition-specific differences over time.

Results
Of the identified significantly expressed genes (Padj &gt; 10−5), five are also implicated as psoriasis risk genes, i.e. HLA-B, SOCS1, MARS, HLA-C and RUNX3. Specifically, a significant increase in RUNX3 expression in wildtype (WT) vs. mutant (MuT) at 16–24 h, and in suppressor of cytokine signalling (SOCS)-1 at 2–24 h was observed, with runt-related transcription factor (RUNX)3 expression peaking at 4 h and SOCS1 showing consistently high expression beyond 0 h. Decreased expression of SOCS1 disrupts IL-23 production and signalling, whereas increased expression of RUNX3 increases T helper (Th)17 and Th22 levels, key cells downstream of IL-23 signalling

Conclusions
Here, we show that carriage of the WT vs. MuT IL23R has downstream effects on expression of HLA-B, SOCS1, MARS, HLA-C and RUNX3, implicating these genes in the IL-23 pathway. This is important as defining pathways that are perturbed in patients has great potential within stratified medicine, with the possibility for better annotation of personalized genetic risk scores.
P34 Evaluating the adaptation of the amputee skin owing to prosthesis use: a study using in vivo Raman spectroscopy
Authors: Hayes Jack , Andrews Jennifer , Andriuskevicius Tomas , Abdelwahab Omar , Briggs Tom , Worsley Peter , Higgins Claire , Masen Marc 
Published: May 2024

Abstract

Introduction and aims
Lower limb prosthesis has proven to be an extremely successful intervention for amputees. However, to achieve adequate function, the device places significant friction and load on the skin, which leads to substantial skin breakdown. Current rehabilitation practices require skin to be ‘trained’ to perform a load-bearing role. Despite this, there are very high injury statistics for skin on the residual limb in the literature, we hypothesize that rehabilitation practices to adapt amputee skin are limited.

Methods
To interrogate this, we used in vivo Raman spectroscopy to evaluate the lower limbs of 13 unilateral lower limb amputees. This enabled us to assess how key constituents of the stratum corneum (SC) such as natural moisturizing factor (NMF), water and lipid content vary with depth. We also analysed sebum content to quantify cytokines present and performed mechanical characterization of the lower limb to investigate stiffness and friction.

Results
We found that the Raman spectra of the amputated limb show reduced NMF, lipid content and water content at the top of the SC when compared with the intact limb (P &lt; 0.05). In addition, we found that amputee skin contains higher levels of inflammatory cytokines compared with control skin. We also recorded that skin on the residuum is four times softer than the intact limb, while friction is much higher at 0.5 on the residuum skin, compared with 0.3 on the intact limb.

Conclusions
Here, we show the adaptations of amputee skin in response to rehabilitation to be minimal. Contrary to the dogma that rehabilitation trains skin to bear load, we found that skin barrier function in amputee skin is impaired, as indicated by altered water and lipid profile in addition to higher baseline inflammation.
Comparative plasma biochemistry analyte data in nesting leatherback (Dermochelys coriacea), foraging green (Chelonia mydas) and foraging and nesting hawksbill (Eretmochelys imbricata) sea turtles in Grenada, West Indies
Authors: Kucinick Madison , Charles Kate E , Carter Kenrith , Edwards Jonnel , Costlow Catherine , Wilkerson Melinda , Seddon Dawn , Marancik David 
Published: May 2024

AbstractBlood biochemistry represents a minimally invasive tool for monitoring sea turtle health, assessing injured sea turtles and supporting conservation strategies. In Grenada, West Indies, plasma biochemical variables were examined in 33 nesting leatherback (Dermochelys coriacea), 49 foraging green (Chelonia mydas), 49 foraging hawksbill (Eretmochelys imbricata) and 12 nesting hawksbill sea turtles sampled between 2017 and 2022. Plasma biochemistry reference intervals are described herein except for nesting hawksbills, which are represented by descriptive statistics due to the low sample size. Select analyte concentrations were positively correlated with curved carapace length in leatherbacks (chloride), green turtles (total protein, albumin and globulin) and foraging hawksbills (total protein, albumin and phosphorus). Cholesterol (7.8 mmol/l ± 1.6 SD) and triglyceride (6.9 mmol/l ± 1.9 SD) concentrations were significantly higher in leatherbacks compared to foraging green turtles, foraging hawksbills and nesting hawksbills (P &lt; 0.001 for all). Cholesterol was significantly higher in nesting hawksbills compared to foraging green turtles (P = 0.050) and foraging hawksbills (P = 0.050). Foraging hawksbills demonstrated significantly higher aspartate transaminase activities than leatherbacks (P = 0.002), green turtles (P = 0.009) and nesting hawksbills (P &lt; 0.001). Biochemical results provide baseline population health data and support guidance for treatments during clinical sea turtle rehabilitation efforts. They also provide insight into species-specific physiologic differences and preludes further studies to better characterize the impacts of life-stage class on biochemistry reference intervals to better support wild sea turtle populations in Grenada.
Associations Between Brain Morphology, Inflammatory Markers, and Symptoms of Fatigue, Depression, or Anxiety in Active and Remitted Crohn’s Disease
Authors: Thomann Anne K , Schmitgen Mike M , Stephan Jule C , Ebert Matthias P , Thomann Philipp A , Szabo Kristina , Reindl Wolfgang , Wolf R Christian 
Published: May 2024

Abstract

Background
Fatigue and psychosocial impairments are highly prevalent in IBD, particularly during active disease. Disturbed brain-gut interactions may contribute to these symptoms. This study examined associations between brain structure, faecal calprotectin, and symptoms of fatigue, depression, and anxiety in persons with Crohn’s disease [CD] in different disease states.

Methods
In this prospective observational study, n = 109 participants [n = 67 persons with CD, n = 42 healthy controls] underwent cranial magnetic resonance imaging, provided stool samples for analysis of faecal calprotectin, and completed questionnaires to assess symptoms of fatigue, depression, and anxiety. We analysed differences in grey matter volume [GMV] between patients and controls, and associations between regional GMV alterations, neuropsychiatric symptoms, and faecal calprotectin.

Results
Symptoms of fatigue, depression, and anxiety were increased in patients with CD compared with controls, with highest scores in active CD. Patients exhibited regionally reduced GMV in cortical and subcortical sensorimotor regions, occipitotemporal and medial frontal areas. Regional GMV differences showed a significant negative association with fatigue, but not with depression or anxiety. Subgroup analyses revealed symptom-GMV associations for fatigue in remitted but not in active CD, whereas fatigue was positively associated with faecal calprotectin in active but not in remitted disease.

Conclusion
Our findings support disturbed brain-gut interactions in CD which may be particularly relevant for fatigue during remitted disease. Reduced GMV in the precentral gyrus and other sensorimotor areas could reflect key contributions to fatigue pathophysiology in CD. A sensorimotor model of fatigue in CD could also pave the way for novel treatment approaches.