bluuebunny/crossref_metadata_2025_split huggingface dataset:
P18 Investigating the role of the microbiome in chronic wound skin barrier reformation
Authors: Cannon Alice , Wilkinson Holly , Rudden Michelle , Hardman Mat 
Published: May 2024

Abstract

Introduction and aims
The integrity of the skin barrier plays a pivotal role in dermatological health, particularly in elderly and individuals with diabetes who are more susceptible to injury. Our skin barrier is also in direct contact with the microbiota, a diverse community of microorganisms that play wide-ranging roles in cutaneous function. Although microbial dysbiosis has been observed in elderly and diabetic skin, few studies have investigated the mechanistic links between skin barrier perturbations and the microbiome. The aim of this study was to compare barrier reformation in healthy and pathological (chronic wound) skin and determine the role of microbial dysbiosis in skin barrier perturbations.

Methods
Here, we developed a novel human ex vivo skin barrier model to simulate skin barrier disruption in healthy and pathological skin. Following tape stripping (n = 6 per donor type), skin barrier disruption was confirmed by measuring transepidermal water loss (TEWL). Skin barrier reformation was assessed following culture using haematoxylin and eosin staining, Nile red (lipid expression and distribution), and immunohistochemistry for skin barrier proteins (e.g. claudins, keratins). Secreted products from mixed bacterial communities were also applied to healthy tape-stripped skin to assess the influence of a healthy vs. pathological microbiome on skin barrier reformation.

Results
Pathological skin showed higher TEWL following barrier disruption, in addition to delayed barrier reformation and altered skin barrier protein expression. Moreover, secreted products from a pathological microbiome, with high proportions of Staphylococcus aureus, significantly delayed skin barrier reformation.

Conclusions
Collectively, our data provide a novel skin barrier reformation model to assess the effect of cutaneous pathology and microbial dysbiosis on skin barrier repair. Further understanding of the role of the microbiome in skin barrier function is integral to reducing infections and barrier perturbation.
P27 Chloride intracellular channel 4 is a global regulator of type 1 interferon signalling in systemic sclerosis skin
Authors: Wasson Christopher , Dibb Sophie , Bryon Jessica , Caballero-Ruiz Begoña , Clavane Eva , Wells Rebecca , Kakkar Vishal , De Lorenzis Enrico , Ross Rebecca , Meakin Paul , Del Galdo Francesco 
Published: May 2024

Abstract

Introduction and aims
Systemic sclerosis (SSc) is an autoimmune disease in which an immune-related injury induces fibrosis of the skin and can progress to affect the internal organs in the most serve cases. Aberrant type 1 interferon (IFN) signalling in the keratinocytes plays a major role in SSc disease progression. We have previously shown the chloride intracellular channel 4 (CLIC)4 is upregulated in SSc skin fibroblasts and plays an important role in SSc fibrosis by enhancing transforming growth factor-β signalling. In this study, we investigated the role of CLIC4 in SSc keratinocyte biology.

Methods
Skin biopsies from patients with SSc (N = 6) and healthy controls (HCs) (N = 6) were analysed by immunohistochemistry for the expression of CLIC4 and phosphorylated signal transducer and activator of transcription (STAT)1. The skin keratinocyte cell line HaCaTs was stimulated with a range of type 1 IFN signalling agonists including POLY I:C, POLY dA:dT, ODN 2216 and IFN-α. CLIC4 was inhibited with the chloride channel inhibitors NPPB and IAA-94 or small interfering RNA specific to CLIC4. Conditioned media from HC or SSc dermal fibroblasts was employed for indirect coculture of HaCaTs.

Results
Skin biopsies from SSc showed high levels of CLIC4 in SSc skin fibroblasts, keratinocytes and endothelial cells compared with HC. Indirect coculture of HaCaTs with SSc fibroblast media induced CLIC4 expression in the HaCaTs. The increased CLIC4 expression played an important role in type 1 IFN signalling in keratinocytes as inhibition of CLIC4 blocked Toll-like receptor (TLR)3, TLR9 and cGAS-mediated activation of the type 1 IFN signalling pathway transcription factor STAT1. Additionally, inhibition of CLIC4 prevented SSc fibroblast media from inducing a type 1 IFN response in keratinocytes.

Conclusions
The data presented in this study suggests that CLIC4 is a global regulator of the type 1 IFN signalling cascade in SSc epithelial cells.
P28 Data-independent acquisition mass spectrometry improves spatially resolved analysis of the human skin proteome
Authors: Ozorun Gulsev , Eckersley Alexander , Bradley Eleanor , Watson Rachel , Sherrat Michael , Swift Joe 
Published: May 2024

Abstract

Introduction and aims
Proteomic analysis of the extracellular matrix (ECM) presents challenges because of the highly crosslinked and low-solubility nature of ECM proteins. Traditional homogenization and protein digestion approaches result in the loss of crucial information regarding protein localization and spatial relationships. To address this, spatially resolved proteomics emerges as a powerful tool for exploring heterogeneity within bulk tissues. This study aims to determine the minimum tissue volume required for comprehensive proteome coverage using data-independent acquisition mass spectrometry (DIA-MS) on skin tissue. The study focused on optimizing spatially resolved proteomic techniques to enhance depth-of-analysis while preserving spatial specificity.

Methods
Human abdominal skin biopsies were obtained from a single individual and subsequently cryosectioned. Histological assessment was performed through haematoxylin and eosin staining for visualization purposes. Laser-capture microdissection coupled with mass spectrometry facilitated the precise isolation of target regions. Comparative analyses were performed between data-dependent acquisition mass spectrometry (DDA-MS) and DIA-MS, with a particular emphasis on ECM proteins within the dermis.

Results
Our findings revealed an improvement in proteome coverage with DIA-MS compared with DDA-MS, in addition to clear scaling relationships between the depth-of-analysis and sample concentration. The Results demonstrated the superiority of DIA-MS in achieving robust and comprehensive proteomic profiles, even with minimal tissue volumes. Preliminary findings suggest the capability of DIA-MS in elucidating the complexities of the skin proteome with spatial resolution.

Conclusions
In conclusion, our study highlights the efficacy of DIA-MS in spatially resolved proteomics on skin tissue. The optimized approach presented here offers a reliable and efficient method for obtaining in-depth proteome information with minimal tissue requirements. These Results form the foundation for ongoing experiments, utilizing DIA-MS to advance spatially resolved proteomic analyses of human skin.
P31 Modelling epidermal melanocytes behaviour during plaque psoriasis resolution
Authors: Lussoso Sarah , Kirby Brian , Tobin Desmond 
Published: May 2024

Abstract

Introduction and aims
Melanocytes in the human epidermis exist in the stratum basale at a ratio of 1 melanocyte to approximately 10 keratinocytes. They do not divide, even upon ultraviolet radiation stimulation, and do not appear to apoptose. In psoriasis, this melanocyte/keratinocyte ratio is retained, despite the enormous keratinocyte hyperplasia, inferring a commensurate expansion in melanocyte number. Therefore, here we investigate whether melanocyte number is altered following the reduction of keratinocytes upon treatment of psoriasis plaques with biologic therapy.

Methods
Matching lesional and nonlesional epidermis sets of patients (n = 4) at day 0 and after 4 and 12 weeks of treatment were compared with healthy controls. To assess the presence of apoptotic melanocytes, the terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assay was performed. Immunohistochemistry with microphthalmia-associated transcription factor (melanocyte marker) and Ki-67 (proliferation marker) was used to assess melanocyte proliferation. Finally, melanocyte number per mm of basal layer and melanocyte/keratinocyte ratio were estimated by image analysis using ImageJ.

Results
Proliferating melanocytes were detected in nonlesional and lesional samples and in resolving plaques after 4 and 12 weeks of treatment but not in healthy epidermis. Melanocyte number per mm of basal layer was significantly higher in nonlesional than lesional sections compared with healthy controls (P &lt; 0.01) suggesting that melanocyte expansion already occurs prior to lesion establishment, revealing early melanocyte deregulation. After 4 and 12 weeks of treatment the number of melanocytes in lesional and nonlesional sections was still significantly higher than controls (P &lt; 0.05) and had not returned to normal despite the dramatic reduction in keratinocytes. The TUNEL assay did not detect evidence of melanocyte apoptosis.

Conclusions
This study shows that rare melanocyte proliferation occurs in nonlesional psoriasis skin and in resolving lesional skin. Given our inability to detect apoptotic melanocytes, it remains unclear how melanocyte numbers return to normal after plaque resolution These unique findings support a role for melanocytes in psoriasis pathogenesis and encourage further investigation.
P35 Adaptation of a phenotypic screening assay to incorporate 384-well plates: opening the door to high-throughput screening
Authors: Feltham Kieran , Lapthorn Alice , Ilg Marcus , Dziewulski Peter , Cellek Selim 
Published: May 2024

Abstract

Introduction and aims
High-throughput screening has significantly impacted drug discovery by increasing speed of experiments and reducing reagent and sample costs. We have previously developed several screening assays to measure myofibroblast inhibition via quantification of α-smooth muscle actin (α-SMA) expression, in a 96-well format. Currently, there are no drugs for the treatment of hypertrophic scarring, a major side-effect of burn injuries that occurs by myofibroblast transformation, leaving a huge gap in this sector. The aim for this project was to adapt our current assay for use in 384-well microplates.

Methods
Human primary dermal fibroblasts were transformed to myofibroblasts using 10 ng mL−1 transforming growth factor (TGF)-β1. α-SMA expression was measured using the In-Cell enzyme-linked immunosorbent assay method, with cell viability measured using DRAQ5 nuclear staining. Assay accuracy and reliability was determined by calculating the robust z-factor, with guidelines set to an average score over 0.5, with a minimum score per plate above 0.4. To further validate the assay, concentration-response curves (CRCs) for the drug vehicle [dimethyl sulfoxide (DMSO)] and our tool compound (SB-505124) were constructed at varying concentrations, in coincubation with TGF-β1.

Results
Optimization of well metrics was performed, producing an average robust z-factor of 0.52, confirming the assay Results are reliable and reproducible. The DMSO CRC produced no significant drop in cell viability at concentrations that are expected to be used in the screening of compounds. Our tool compound (SB-505124) produced a concentration-dependant curve for the inhibition of myofibroblast transformation.

Conclusions
A 384-well assay has been designed, tested and validated for the determination of myofibroblast transformation in human primary dermal fibroblasts. The assay has been shown to deliver an average robust z-factor of above 0.5, it contains a positive control and is compatible with compounds stored in up to 100% DMSO.
P37 Drug repurposing for treatment of recessive dystrophic epidermolysis bullosa cutaneous squamous cell carcinoma
Authors: Pritchard John , Dayal Jasbani , Schoenherr Christina , Raponi Michela , Anand Jayanthi , McGarry Lynn , Corbyn Ryan , Carlin Leo , Blyth Karen , Inman Gareth 
Published: May 2024

Abstract

Background
Recessive dystrophic epidermolysis bullosa (RDEB) is an inherited skin disorder that leads to severe wounding and blistering of the skin. Patients with RDEB develop aggressive cutaneous squamous cell carcinoma (cSCC) with a mortality rate of approximately 90% by age 55 years. The current primary treatment is wide excision of tumours, often leading to amputations. Identifying new therapeutic avenues for RDEB cSCC remains a priority. Here, we approach this in two ways. Firstly, we identified a SMAD4-dependent sphingosine pathway response to endogenous transforming growth factor (TGF)-β, essential for RDEB cSCC proliferation which could be targeted as a potential therapeutic strategy. Secondly, we utilized a US Food and Drug Administration (FDA) screen of &gt; 3000 drugs to identify potential druggable pathways that could be used to treat RDEB cSCC.

Methods
RDEB cSCC cell lines were used to test Siponimod, a sphingosine-1 receptor modulator, on proliferation, clonogenicity and invasion in vitro. Siponimod was used in vivo to test tumour growth effect of targeting the sphingosine pathway. We carried out an FDA-approved drug screen on four RDEB cSCC cell lines using the Opera Phenix high-throughput microscope to identify targets.

Results
Treatment of RDEB cSCC cell lines with Siponimod showed compromised proliferation compared with untreated cells in addition to significant reduction in colony formation. There was a significant decrease in the invasive capability of RDEB SCC cells. Treatment of normal human keratinocytes showed no effect on proliferation. Our FDA-approved screen identified multiple drugs that target MEK, mechanistic target of rapamycin complex, histone deacetylase, signal transducer and activator of transcription, and topoisomerase I/II in the top 50 hits. The most common target was DNA damage/repair pathways, but cytoskeletal signalling, transmembrane transporters and proteases were identified as other potential avenues for treatment of RDEB cSCC.

Conclusions
There is an unmet need for therapeutic options in RDEB cSCC. Here, we have demonstrated the potential of repurposing previously licensed drugs to identify treatment avenues for RDEB cSCC.
Overall survival after mastectomy versus breast-conserving surgery with adjuvant radiotherapy for early-stage breast cancer: meta-analysis
Authors: Rajan Kiran K , Fairhurst Katherine , Birkbeck Beth , Novintan Shonnelly , Wilson Rebecca , Savović Jelena , Holcombe Chris , Potter Shelley 
Published: May 2024

Abstract

Background
Breast-conserving surgery with adjuvant radiotherapy and mastectomy are currently offered as equivalent surgical options for early-stage breast cancer based on RCTs from the 1970s and 1980s. However, the treatment of breast cancer has evolved and recent observational studies suggest a survival advantage for breast-conserving surgery with adjuvant radiotherapy. A systematic review and meta-analysis was undertaken to summarize the contemporary evidence regarding survival after breast-conserving surgery with adjuvant radiotherapy versus mastectomy for women with early-stage breast cancer.

Methods
A systematic search of MEDLINE, the Cochrane Central Register of Controlled Trials (CENTRAL), and Embase that identified studies published between 1 January 2000 and 18 December 2023 comparing overall survival after breast-conserving surgery with adjuvant radiotherapy versus mastectomy for patients with unilateral stage 1–3 breast cancer was undertaken. The main exclusion criteria were studies evaluating neoadjuvant chemotherapy, rare breast cancer subtypes, and specific breast cancer populations. The ROBINS-I tool was used to assess risk of bias, with the overall certainty of evidence assessed using the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) tool. Studies without critical risk of bias were included in a quantitative meta-analysis.

Results
From 11 750 abstracts, 108 eligible articles were identified, with one article including two studies; 29 studies were excluded from the meta-analysis due to an overall critical risk of bias, 42 studies were excluded due to overlapping study populations, and three studies were excluded due to reporting incompatible results. A total of 35 observational studies reported survival outcomes for 909 077 patients (362 390 patients undergoing mastectomy and 546 687 patients undergoing breast-conserving surgery with adjuvant radiotherapy). The pooled HR was 0.72 (95% c.i. 0.68 to 0.75, P &lt; 0.001), demonstrating improved overall survival for patients undergoing breast-conserving surgery with adjuvant radiotherapy. The overall certainty of the evidence was very low.

Conclusion
This meta-analysis provides evidence suggesting a survival advantage for women undergoing breast-conserving surgery with adjuvant radiotherapy for early-stage breast cancer compared with mastectomy. Although these results should be interpreted with caution, they should be shared with patients to support informed surgical decision-making.
Older Adult Homicide: Investigating Case, Victim and Perpetrator Characteristics in a National Sample from England and Wales
Authors: Storey Jennifer E , Rogers Michaela M , Hohn Richard E 
Published: May 2024

Abstract
Older adult homicide (OAH) is the most severe, yet understudied, form of older adult abuse. This study examined the case, victim and perpetrator characteristics of OAH. A secondary analysis of national data from England and Wales (2008–2019) was conducted where cases of non-stranger OAH (victims aged sixty years and over) were compared to adult homicide (victims aged eighteen to fifty-nine years) at the case, victim (n = 3,274) and perpetrator (n = 2,763) levels. Logistic regression models used to identify characteristics that were OAH risk factors, showed only a slight increase in predictive power but high accuracy in classifying adult homicide cases. Nevertheless, some risk factors known to be predictors of older adult abuse were significant predictors of OAH (e.g. living with the perpetrator, the perpetrator’s mental state). Implications for research, policy and practice are discussed.
Effects of evening smartphone use on sleep and declarative memory consolidation in male adolescents and young adults
Authors: Höhn Christopher , Hahn Michael A , Gruber Georg , Pletzer Belinda , Cajochen Christian , Hoedlmoser Kerstin 
Published: May 2024

Abstract
Exposure to short-wavelength light before bedtime is known to disrupt nocturnal melatonin secretion and can impair subsequent sleep. However, while it has been demonstrated that older adults are less affected by short-wavelength light, there is limited research exploring differences between adolescents and young adults. Furthermore, it remains unclear whether the effects of evening short-wavelength light on sleep architecture extend to sleep-related processes, such as declarative memory consolidation. Here, we recorded polysomnography from 33 male adolescents (15.42 ± 0.97 years) and 35 male young adults (21.51 ± 2.06 years) in a within-subject design during three different nights to investigate the impact of reading for 90 min either on a smartphone with or without a blue-light filter or from a printed book. We measured subjective sleepiness, melatonin secretion, sleep physiology and sleep-dependent memory consolidation. While subjective sleepiness remained unaffected, we observed a significant melatonin attenuation effect in both age groups immediately after reading on the smartphone without a blue-light filter. Interestingly, adolescents fully recovered from the melatonin attenuation in the following 50 min before bedtime, whereas adults still, at bedtime, exhibited significantly reduced melatonin levels. Sleep-dependent memory consolidation and the coupling between sleep spindles and slow oscillations were not affected by short-wavelength light in both age groups. Nevertheless, adults showed a reduction in N3 sleep during the first night quarter. In summary, avoiding smartphone use in the last hour before bedtime is advisable for adolescents and young adults to prevent sleep disturbances. Our research empirically supports general sleep hygiene advice and can inform future recommendations regarding the use of smartphones and other screen-based devices before bedtime.
Palladium-iron bimetallic-catalyzed cross-coupling reaction of alcohols and nitroarenes using a hydrogen-transfer redox system
Authors: Horikawa Rikiya , Ishii Shoma , Shinozaki Urara , Midorikawa Reika , Mori Takamichi , Kimura Masanari 
Published: May 2024

Abstract
A Pd/Fe bimetallic catalyst in the presence of the Xantphos ligand promoted the cross-coupling reaction of alcohols and nitroarenes via a hydrogen-transfer redox system. The consecutive formation of aldimines from benzyl alcohols and nitroarenes proceeded in three steps in a single manipulation: oxidation of alcohols, reduction of nitroarenes, and dehydrative condensation of aldehydes and aminoarenes.
CMR characterization of patients with heart failure and left bundle branch block
Authors: Tomoaia Raluca , Harrison Peter , Bevis Lydia , Wahab Ali , Thompson Patrick , Saunderson Christopher E D , Levelt Eylem , Dall’Armellina Erica , Garg Pankaj , Greenwood John P , Plein Sven , Swoboda Peter P 
Published: May 2024

Abstract

Aims
We aimed to identify the distinctive cardiovascular magnetic resonance (CMR) features of patients with left bundle branch block (LBBB) and heart failure with reduced ejection fraction (HFrEF) of presumed non-ischaemic aetiology. The secondary aim was to determine whether these individuals exhibit characteristics that could potentially serve as predictors of left ventricular ejection fraction (LVEF) recovery as compared with patients without LBBB.

Methods and results
We prospectively recruited patients with HFrEF (LVEF ≤ 40%) on echocardiography who were referred for early CMR examination. Patients with an established diagnosis of coronary artery disease and known structural or congenital heart disease were excluded. LV recovery was defined as achieving ≥10% absolute improvement to ≥40% in LVEF between baseline evaluation to CMR. A total of 391 patients were recruited including 115 (29.4%) with LBBB. Compared with HF patients without LBBB, those with LBBB exhibited larger left ventricles and smaller right ventricles, but no differences were observed with respect to LVEF (35.8 ± 12 vs. 38 ± 12%, P = 0.105). The overall rate of LV recovery from baseline echocardiogram to CMR (70 [42–128] days) was not significantly different between LBBB and non-LBBB patients (27.8% vs. 31.5%, P = 0.47). Reduced LVEF remained an independent predictor of LV non-recovery only in patients with LBBB.

Conclusion
Patients presenting with HFrEF and LBBB had larger LV cavities and smaller RV cavities than those without LBBB but no difference in prevalence of scar or ischaemia. The rates of LV recovery were similar between both groups, which supports current guidelines to defer device therapy until 3–6 months of guideline-directed medical therapy, rather than early CMR and device implantation.