bluuebunny/crossref_metadata_2025 huggingface dataset:
High
β
magnetic reconnection at different altitudes in the cool low solar atmosphere
Authors: Zafar Abdullah , Ni Lei , Lin Jun , Ziegler Udo
Published: July 2023

We numerically studied magnetic reconnection in a high β hydrogen–helium plasma at different altitudes from the photosphere to the upper chromosphere. The time-dependent ionization degrees were included to get more realistic diffusivities and viscosity, and appropriate radiative cooling models were applied. Our numerical results indicate that the plasmoid instability always plays a vital role in speeding up magnetic reconnection at different atmospheric layers. In addition, both the strong radiative cooling and the magnetic diffusion caused by the electron–neutral collision (ηen) can significantly accelerate magnetic reconnection below the middle chromosphere. On the other hand, both the ambipolar diffusion and the viscosity result in higher temperature and plasma pressure in the reconnection region in the upper chromosphere, which then hinders the fast reconnection process from developing. The local compression heating triggered by turbulent reconnection mediated with plasmoids is the dominant heating mechanism in the unstable reconnection stage at different atmospheric layers, but the viscous heating and the ambipolar diffusion heating are equally important in the upper chromosphere. The Joule heating contributed by ηen dominates during the early quasi-steady reconnection stage below the middle chromosphere, the strong radiative cooling also leads to much stronger compression heating and more generation of thermal energy in this region. Though the plasma β is the same in all the simulation cases at different altitudes, the temperature increase is more significant in the upper chromosphere with much lower density and weaker radiative cooling.
Theory and effectiveness analysis of dual-aircraft formation active cancellation slow synchronous blinking jamming
Authors: Hao Zelong , Wang Xing , Chen You
Published: July 2023

Dual-aircraft formation slow synchronous blinking jamming is standard for jamming monopulse tracking. However, the jamming signal ratio (JSR) of slow synchronous blinking jamming is low and fluctuant in a dynamic jamming scenario, which leads to a poor jamming effect. This study improves the theory of active cancellation slow synchronous blinking jamming (ACBJ) by combining the cancellation stealth technology with slow synchronous blinking jamming to enhance the slow synchronous blinking jamming. We develop a motion model of dual-aircraft formation in the dynamic jamming scenario. In addition, the dynamic radar cross-section for stealth and nonstealth aircraft is simulated separately. The ACBJ theoretical model and its jamming effectiveness are then derived. We also propose the whole process jamming effectiveness (WPJE) index, and a local-global evaluation index is constructed based on WPJE to evaluate jamming effectiveness reasonably. Finally, we designed four experiments to prove that ACBJ can increase the JSR and improve the jamming effect. The WPJE index for nonstealth aircraft formation using ACBJ was 24% higher than that for slow synchronous blinking jamming, and the WPJE index for stealth aircraft formation using ACBJ increased from 98% to 100%.
An infrared spectroscopic study on gaseous molecular clusters: (Acrylonitrile–methanethiol)+ and (acrylonitrile–dimethyl sulfide)+
Authors: Xu Yingbo , Zhang Jiayang , Xie Min , Hu Yongjun
Published: July 2023

The ion–molecule reaction is one of the most important pathways for the formation of new interstellar chemical species. Herein, infrared spectra of cationic binary clusters of acrylonitrile (AN) with methanethiol (CH3SH) and dimethyl sulfide (CH3SCH3) are measured and compared to those previous studies of AN and methanol (CH3OH) or dimethyl ether (CH3OCH3). The results suggest that the ion–molecular reactions of AN with CH3SH and CH3SCH3 only yield products with S…HN H-bonded or S∴N hemibond structures, rather than the cyclic products as observed in AN-CH3OH and AN-CH3OCH3 studied previously. The Michael addition-cyclization reaction between acrylonitrile and sulfur-containing molecules does not occur due to the weaker acidity of CH bonds in sulfur-containing molecules, which results from their weaker hyperconjugation effect compared to oxygen-containing molecules. The reduced propensity for the proton transfer from the CH bonds hinders the formation of the Michael addition-cyclization product that follows.
The effect of single-doping engineering on ferroelectric properties of α-In2Se2X: A density functional theory study
Authors: Wen Jun , Chen Yi-Jie , Sun Fu-Li , Lin Cun-Biao , Zhang Xian-Jie , Wang Jia-Hao , Chen Wen-Xian , Zhuang Gui-Lin
Published: July 2023

Tailoring the specific properties to practical applications by structural modification is of vital importance for the envisioned development of two-dimensional ferroelectric materials. Herein, a comprehensive investigation on the effects of single doping on the ferroelectric properties and electronic transport in a monolayer of α-In2Se3 was carried out via the combination of first-principles density functional theory calculations and electron–phonon coupling simulations. Our results show that single-doping in In2Se3 can reduce effective mass of carriers and thereby enhance the high carrier mobility potential of the material. Moreover, the ferroelectric phonon mode in single-doped In2Se2X features a lower scattering rate, associating with the single-doping atom, and indicates reduced hindrance to carrier transport during ferroelectric switching. Compared to pristine In2Se3, the obtained smaller ferroelectric barriers (<1 eV) of single-doped ones promote more promising ferroelectricity from the analysis of the ferroelectric soft mode. Interestingly, the observed variations in ferroelectric behaviors resulting from doping of different elements highlight the significance of single-doping in modifying the ferroelectric properties of monolayers. Furthermore, strain engineering results reveal that single doping obviously affects the dependence of gap on strains: linear relationship for doping ones and nonlinearity for pristine one. Our study provides valuable insights into achieving higher carrier mobility in these critical materials.
Absorption limit in direct gap III–V semiconductors
Authors: Bhowmick Mithun , Xi Haowen , Ullrich Bruno
Published: July 2023

The comparison of experimentally found absorption limits, and their variations in compound semiconductors, with theoretical expectations was never a particularly studied subject, although absorption limits in semiconductors could be critical to certain optoelectronic device applications. We introduce a model, which accurately fits the distinct absorption saturation parameter linked to the effective electron density of states.
The role of primary care in optimising women’s sexual and reproductive health
Authors: Mazza Danielle , Botfield Jessica R.
Published: July 2023

The papers in this special issue explore a range of issues and opportunities across the spectrum of sexual and reproductive health in primary care, including in relation to contraception access; support for unintended pregnancies; abortion care; preconception, pregnancy, postpartum and interconception care; and issues relating to informed consent. The special issue takes a reproductive life course perspective through consideration of both pregnancy prevention and pregnancy preparation.
Informational communication and metacognition
Authors: Proust Joëlle
Published: July 2023

Procedural metacognition is the set of affect-based mechanisms allowing agents to regulate cognitive actions like perceptual discrimination, memory retrieval or problem solving. This article proposes that procedural metacognition has had a major role in the evolution of communication. A plausible hypothesis is that, under pressure for maximizing signalling efficiency, the metacognitive abilities used by nonhumans to regulate their perception and their memory have been re-used to regulate their communication. On this view, detecting one’s production errors in signalling, or solving species-specific trade-offs between informativeness, processing effort, clarity, or urgency depend on a form of procedural metacognition, called “metacommunication”. How does this view relate to Gricean theories of human communication? A parallel between procedural trade-offs and conversational maxims is discussed for its evolutionary implications. Rather than accepting radically discontinuist interpretations, in which mindreading operates a full reorganization of pragmatics, it is proposed that procedural forms of regulation are entrenched in all forms of human communication. According to contextual demands, humans adopt and monitor more or less demanding informational goals, such as factual updating, clarifying, explaining, proving, and reaching consensus in collective matters. Under time pressure, only part of these goals require adopting others’ viewpoint. Efficiency in collective decision-making, in particular, might have been considerably raised by an ability to interpret others’ intentions and motivations.
Rapid evaluation of the particle-erosion resistance of Al2O3 ceramics, composites, and coatings using a resonant acoustic mixer
Authors: Mechnich Peter , Alkan Gözde
Published: July 2023

The solid particle technology usage in concentrated solar power plants as direct heat absorption and storage medium necessitate well selection of the materials for the components such as transport and sluice systems, which are in direct contact with moving and falling hot particles up to 1500 ˚C. Beyond mechanical properties, chemical inertness and high-temperature stability, abrasion/erosion resistance are one of the key properties, for which, there is no easy-applicable and rapid test method exist enabling controlled lab-scale parametric studies. A novel particle impact test was established using a resonance acoustic mixer, in which ceramic particles are strongly accelerated and collide with the ceramic surface within a closed vessel. After determination of the most representative parameters such as ceramic ball size, vessel diameter, and retainment/removal of debris, selected experiments were conducted on three candidate materials aimed to be used as high-temperature transport/port systems; dense C 799 Al2O3, porous water-plasma sprayed Plascera-type Al2O3 and WHIPOX-type Al2O3/Al2O3 ceramic matrix composites with porous matrix; with and without porous protective Al2O3 coating. The distinct mass loss behaviour of candidate materials highlighted the viability of the test method and the relevance of microstructures of porous Al2O3 materials on abrasion resistance.
Decision-making algorithms for the identification of crops based on remote sensing data
Authors: Sulyukova L F , Shamsiev R Z
Published: July 2023

The article presents an algorithm for collective management decision-making. The developed algorithm is based on Bayesian mechanisms for resolving contradictions when choosing methods for processing of decisions. The algorithm is used for phased processing of Earth remote sensing data in order to identify agricultural crops. Functional satellite images created using the algorithm increase the accuracy of identifying plant objects. With the help of GPS navigator observation points linked by geographical coordinates, reference points have been created, according to which the classification of functional satellite images is accomplished, with the most accurate results of the distribution of pasture crops in contrast to the original image. The classification was performed according to four types of vegetation in the studied territory of Northern and Southern Nuratau of the Republic of Uzbekistan. Based on the results obtained, field work was carried out in order to test the remote sensing data obtained in relation to the identification of agricultural crops of pasture territories.
Application of artificial intelligence technologies to assess water salinity
Authors: Muhamediyeva D T
Published: July 2023

Topical issues of developing theoretical and methodological tools for constructing a fuzzy logical model for assessing water salinity are considered. When constructing a Sugeno fuzzy logical model for estimating water salinity, a rational number of rules and effective values of their membership functions were chosen. Initially, the membership function parameters were obtained from water industry experts. In the future, it is necessary to adjust the parameters of the membership function using neural networks to obtain the minimum number of fuzzy rules.
Factors affecting the highly dispersed spray when applying fertilizers
Authors: Salikhova M N , Aleksandrov V A , Galchak I P , Kolga A D , Kukhar V S
Published: July 2023

The article is devoted to devices for subsurface fertilization in the form of drip-air mixtures. A comparative analysis of spray mixers with centrifugal and jet nozzles has been carried out. The article presents the results of a study on the spray dependence on the ratio of air and liquid pressure with increasing fluid flow and steady air flow. It has been established that a spray mixer with a centrifugal nozzle gives a thinner spray cone is better in its operational qualities and will more fully meet the requirements for devices for intra-soil application of liquid fertilizers. In addition, it was determined that according to agrotechnical requirements, the average diameter of liquid droplets injected into the soil must be kept constant, and the flow rate must be changed, leaving the device speed and the compressor operating mode unchanged.
Rapid synthesis of nickel ferrite nanoparticles by the molten salt method
Authors: León-Flores J , Pérez-Mazariego J L , Olmedo-Reséndiz E T , Marquina M L , Escamilla R , Gómez R , Arenas-Alatorre J
Published: July 2023

It is presented a ‘shocking thermal treatment’ methodology as a modification to the molten salts process to rapidly obtain pure phase nickel ferrite nanoparticles within a high-scalable green synthesis scheme. Analysis from x-ray diffraction and electron microscopy (SEM and TEM) confirms the formation of the nanostructures, with the most frequent crystallite sizes between 55–85 nm. Room temperature magnetic characterization, from Mössbauer spectroscopy and magnetic measurements M(H), evidences their superparamagnetic distribution with a magnetic saturation of 47.9 emu g−1. An approximated stoichiometry of

Ni

0.03

Fe

0.97

A

Ni

0.97

Fe

1.03

B

O

4

was proposed from Rietveld refinement, Mössbauer and energy-dispersive x-ray spectroscopies. The effect on the electronic structure due to the shocking thermal treatment was elucidated by estimating the optical band gap with values between 1.78 and 2.38 eV.
High-efficiency photon-number-resolving detector for improving heralded single-photon sources
Authors: Stasi Lorenzo , Caspar Patrik , Brydges Tiff , Zbinden Hugo , Bussières Félix , Thew Rob
Published: July 2023

Heralded single-photon sources (HSPS) intrinsically suffer from multiphoton emission, leading to a trade-off between the source’s single-photon quality and the heralding rate. A solution to this problem is to use photon-number-resolving (PNR) detectors to filter out the heralding events where more than one photon pair is created. Here, we demonstrate an improvement of a HSPS by heralding photons using a high-efficiency parallel superconducting nanowire single-photon detector (P-SNSPD) with PNR power. Specifically, we show a reduction in the

g

(
2
)

(
0
)

of the heralded single photon by

(
26.9
±
0.1
)
%

for a fixed pump power, or alternatively, an increase in the heralding rate by a factor of

1.368
±
0.002

for a fixed

g

(
2
)

(
0
)

. We also demonstrate that such a PNR device can reveal thermal photon-number statistics of unheralded photons, which is enabled by our ability to construct its full input–output response function. These results are possible thanks to our P-SNSPD architecture that ensures non-latching operation with no electrical crosstalk, which are essential conditions necessary to obtain the correct photon-number statistics and also faster recovery times, therefore enabling fast heralding rates. These results show that our efficient PNR P-SNSPD architecture can significantly improve the performance of HSPSs and can precisely characterize them, making these detectors a useful tool for a wide range of optical quantum information protocols.
3D printing of functional bioengineered constructs for neural regeneration: a review
Authors: Zhu Hui , Yao Cong , Wei Boyuan , Xu Chenyu , Huang Xinxin , Liu Yan , He Jiankang , Zhang Jianning , Li Dichen
Published: July 2023

Highlights

Various 3D printing techniques for neural tissue-engineered scaffolds or living cell-laden constructs are summarized and compared.

Strategies by integrating topographical, biochemical and electroactive cues inside 3D-printed neural constructs for functional neural regeneration were introduced.

The typical applications of 3D-printed bioengineered constructs are demonstrated.

The challenges and future outlook associated with 3D printing for functional neural constructs in various categories are discussed.
Integral cryptanalysis: a new key determination technique for 3-phase Kuznyechik encryption
Authors: Ilkhom Boykuziev , Khan Angshuman , Bakhtiyor Abdurakhimov , Das Rupayan , Khudoykulov Zarif
Published: July 2023

The primary objective of this work is to develop an algorithm for detecting the secret round key used in encryption by analyzing a collection of plaintexts and ciphertexts encrypted with the three-round Kuznyechik algorithm. In addition, the fundamentals of using the integral cryptanalysis method for the Substitution-permutation (SP) network-based Kuznyechik encryption have been investigated. It has been found that the Kuznyechik three-round encryption algorithm is useful for creating a balanced set before the third round and an unbalanced set at the end of the round, using a set in which the first byte is active, and the remaining bytes are passive. Based on the findings above, an efficient algorithm to determine the key of the final round for the 3-round Kuznyechik algorithm is proposed utilizing the method of integral cryptanalysis. According to simulation results, the proposed approach offers approximately 90% improvement compared to baseline algorithms in terms of time complexity, data, and memory overhead.
Buccal Fat Pad Augmentation for Midfacial Rejuvenation: Modified Fat Grafting Technique and Ogee Line Remodeling
Authors: Zhu Congxiao , Li Facheng , Tang Xiaojun , Cai Lei , Yin Bo , Zhang Xinyu , Jiang Chanyuan , Han Xuefeng
Published: July 2023

Background

The volume and position of the buccal fat pad (BFP) change with age, which manifests as a hollow midface. Previous studies showed that autologous fat grafting for BFP augmentation could effectively ameliorate midfacial hollowing.

Objectives

The aim of this study was to introduce a modified fat grafting technique for female patients with midfacial hollowing to restore the volume of BFP, and to evaluate the safety and effectiveness of this approach.

Methods

Two cadavers were used for the dissection of the BFP and to demonstrate the surgical procedures. Forty-eight patients were treated for midfacial hollowing with the modified grafting strategy. The BFP was filled through a percutaneous zygomatic incision and an immediate amelioration in the hollow area was observed. Improvements were evaluated from measurements of the ogee line and ogee angle, FACE-Q questionnaires, and 3-party satisfaction ratings. Clinical profiles were reviewed and statistically analysed.

Results

The mean [standard deviation] ogee angle was 6.6° [1.9°] preoperatively and 3.9° [1.4°] postoperatively (average reduction, 2.7°). Patients’ ogee lines were smoother postoperatively, with marked improvements in overall appearance, psychological well-being, and social confidence. Patients reported high satisfaction with decision-making and postoperative outcomes and felt 6.61 [2.21] years younger. Overall, 88%, 76%, and 83% of the cases were graded as good or excellent in improvement by surgeon, patient, and the third party, respectively.

Conclusions

For age-dependent midfacial hollowing in female patients, the modified percutaneous grafting technique described here was safe and efficacious in restoring BFP volume. This technique produced a smoother ogee line and a natural, younger midfacial contour.

Level of Evidence: 4
Deep multi-omics integration by learning correlation-maximizing representation identifies prognostically stratified cancer subtypes
Authors: Ji Yanrong , Dutta Pratik , Davuluri Ramana
Published: July 2023

Motivation

Molecular subtyping by integrative modeling of multi-omics and clinical data can help the identification of robust and clinically actionable disease subgroups; an essential step in developing precision medicine approaches.

Results

We developed a novel outcome-guided molecular subgrouping framework, called Deep Multi-Omics Integrative Subtyping by Maximizing Correlation (DeepMOIS-MC), for integrative learning from multi-omics data by maximizing correlation between all input -omics views. DeepMOIS-MC consists of two parts: clustering and classification. In the clustering part, the preprocessed high-dimensional multi-omics views are input into two-layer fully connected neural networks. The outputs of individual networks are subjected to Generalized Canonical Correlation Analysis loss to learn the shared representation. Next, the learned representation is filtered by a regression model to select features that are related to a covariate clinical variable, for example, a survival/outcome. The filtered features are used for clustering to determine the optimal cluster assignments. In the classification stage, the original feature matrix of one of the -omics view is scaled and discretized based on equal frequency binning, and then subjected to feature selection using RandomForest. Using these selected features, classification models (for example, XGBoost model) are built to predict the molecular subgroups that were identified at clustering stage. We applied DeepMOIS-MC on lung and liver cancers, using TCGA datasets. In comparative analysis, we found that DeepMOIS-MC outperformed traditional approaches in patient stratification. Finally, we validated the robustness and generalizability of the classification models on independent datasets. We anticipate that the DeepMOIS-MC can be adopted to many multi-omics integrative analyses tasks.

Availability and implementation

Source codes for PyTorch implementation of DGCCA and other DeepMOIS-MC modules are available at GitHub (https://github.com/duttaprat/DeepMOIS-MC).

Supplementary information

Supplementary data are available at Bioinformatics Advances online.
MDSubSampler: a posteriori sampling of important protein conformations from biomolecular simulations
Authors: Oues Namir , Dantu Sarath Chandra , Patel Riktaben Jigarkumar , Pandini Alessandro
Published: July 2023

Motivation

Molecular dynamics (MD) simulations have become routine tools for the study of protein dynamics and function. Thanks to faster GPU-based algorithms, atomistic and coarse-grained simulations are being used to explore biological functions over the microsecond timescale, yielding terabytes of data spanning multiple trajectories, thereby extracting relevant protein conformations without losing important information is often challenging.

Results

We present MDSubSampler, a Python library and toolkit for a posteriori subsampling of data from multiple trajectories. This toolkit provides access to uniform, random, stratified, weighted sampling, and bootstrapping sampling methods. Sampling can be performed under the constraint of preserving the original distribution of relevant geometrical properties. Possible applications include simulations post-processing, noise reduction, and structures selection for ensemble docking.

Availability and implementation

MDSubSampler is freely available at https://github.com/alepandini/MDSubSampler, along with guidance on installation and tutorials on how it can be used.
O02 Neoantigens from actinic keratosis are predicted to be more immunogenic than those from cutaneous squamous cell carcinoma – a strategy for immune escape?
Authors: Thomson Jason , Harwoood Catherine , Strid Jessica  , Healy Eugene , Wang Jun
Published: July 2023

Actinic keratosis (AK) is a precursor of cutaneous squamous cell carcinoma (cSCC). More than 70% of cSCC arise from AKs, but fewer than 0.6% of AKs progress to cSCC. We have shown that the molecular landscapes of AK and cSCC are similar in terms of tumour mutational burden (TMB), copy number burden, mutational signatures and driver gene mutations but differ in levels of intrasample heterogeneity. We have also demonstrated that alterations in signalling pathways, including transforming growth factor-β, may be important for AK-to-SCC progression. However, the pathomechanisms underlying progression are not well characterized and it remains unclear why the majority of AKs do not progress, and most will regress. There is some evidence that differences in the neoantigen landscapes of AK and cSCC may play a role, and a previous study examining neoantigens in AK and cSCC from nine patients was the first to suggest that there is downregulation of the expression of highly immunogenic neoantigens in the immune escape of cSCC. In this study we further explore, in a larger data set, the hypothesis that the neoantigen recognition potential of AK vs. cSCC is a key determinant of AK-to-cSCC progression. We generated exome and transcriptome data from AK (n = 5) and cSCC (n = 15) with matched normal exome data (from blood). Human leucocyte antigen typing was performed using the OptiType algorithm and neoantigens restricted to major histocompatibility complex (MHC) class I were predicted using the NeoPredPipe workflow. Fitness of neoantigens, defined as the overall effectiveness of a neoantigen at stimulating an immune response, was determined by combining measures of MHC binding affinity and T-cell receptor recognition potential for the predicted neoantigen, and an overall neoantigen recognition potential score was derived for each neoantigen. The difference in median total neoantigen burden for AK and cSCC was not statistically significant [11 224 (range 30–17 330) and 1680 (range 3159–67 357), respectively (P = 0.07)]. Neoantigen burden was strongly correlated with TMB in both AK and cSCC. We found that, compared with cSCC, AK had predicted neoantigens with significantly higher mean MHC-binding affinity (P = 0.049), mean T-cell receptor recognition potential (P = 0.0093) and overall neoantigen recognition potential (P = 0.02). Taken together, our results confirm that neoantigens in AKs are predicted to be more effective at eliciting an immune response than those in cSCC. This suggests there may be a selection pressure in AKs for mutations that result in fewer immunogenic neoantigens resulting in immune escape and progression to cSCC. Finally, our results underscore the critical requirement for an intact immune system in prevention of AK-to-cSCC progression.
O11 The human discs large protein (Dlg1) controls connexin 43 (Cx43) plasma membrane localization and gap junctional communication in keratinocytes
Authors: Scott Harry , Dong Li , Srinivasan Sharmila , Stevenson Andrew , MacDonald Alasdair , Massimi Paola  , Banks Lawrence , Martin Patricia , Johnstone Scott , Graham Sheila
Published: July 2023

Gap junctional communication is required for cellular coordination in health and disease. Gap junction channels are composed of connexins and allow direct intercellular ­communication. Connexins also play an important role in processes such as wound healing, with connexin 43 (Cx43) being downregulated at the plasma membrane of cells at the wound edge during the initial stages of the normal wound healing process. Connexins must be located on the plasma membrane in order to form gap junctions; therefore, it is important to identify proteins involved in the regulation of this. Previously, in a study of human papillomavirus (HPV)-positive tumour cells, we reported that the gap junction protein Cx43 is a physiologically relevant binding partner of the human homologue of Drosophila Discs large (Dlg1). Dlg1 is a member of the membrane associated-guanylate kinase (MAGUK) scaffolding protein family that is known to control cell shape and polarity, and intracellular trafficking. Here we show, for the first time, that Cx43 interacts with Dlg1 in uninfected keratinocytes in vitro and in normal human epithelial tissues. Depletion of Dlg1 in keratinocytes did not alter Cx43 transcription but was associated with an overall reduction in Cx43 protein levels. The loss of Dlg1 in keratinocytes was further associated with a loss of Cx43 at the plasma membrane and a reduction in intercellular communication. This decrease in Cx43 levels was partially reversed through use of a lysosomal inhibitor; however, a proteasomal inhibitor resulted in no difference to Cx43 levels, suggesting that, in the absence of Dlg1, Cx43 is degraded through the lysosomal but not the proteasomal pathway. This correlated with an increased association between Cx43 and LAMP2, a marker of lysosomes in Dlg1 knockdown cells. In the absence of Dlg1, Cx43 localized to the Golgi but not the endoplasmic reticulum. Although this result could indicate that Dlg1 is required for stable Cx43 membrane location, a study of Cx43 and Dlg1 during the wound healing process in HaCaT cells (a spontaneously immortalized HPV-negative epithelial cell line), showed the proteins co-localizing as they relocated from the plasma membrane to the cytoplasm between 4 and 16 h postwounding before returning to the plasma membrane at 24 h postwounding. This may suggest that Dlg1 is involved in Cx43 trafficking. The levels of both proteins were reduced between 0 and 4 h postwounding and then increased between 4 and 16 h, before returning to normal levels at 24 h postwounding. Taken together, our data suggest that Dlg1 is crucial for promoting plasma membrane localization of Cx43 in keratinocytes.
O15 Using CRISPR-Cas9 to annotate the TYK2 pathway in psoriasis
Authors: Elyoussfi Sarah , Rane Shraddha S , Eyre Stephen  , Warren Richard B
Published: July 2023

Genome-wide association studies (GWAS) have discovered genetic signals associated with psoriasis. This study is the first to validate functionally the psoriasis-associated single nucleotide polymorphism (SNP) rs892085 on TYK2 expression using clustered regularly interspaced short-palindromic repeat Cas9 endonuclease (CRISPR-Cas9) technologies in Jurkat T-lymphoblast cells. Our in-house capture Hi-C data revealed long-range chromatin interactions between specific SNP sets and TYK2. An independent noncoding genetic association with psoriasis was identified about 400 kb upstream of TYK2: rs892085 at 19p13.2. This is a psoriasis-associated risk SNP (P = 3.0 × 10–17). To characterize fully the physical connection, CRISPR-Cas9 genome editing was conducted in T-cell-like Jurkat T-lymphoblast cell lines. We used the CRISPR activator (CRISPR/dCas9-VP64) and CRISPR interference (CRISPR/dCas9-KRAB) systems to alter the transcriptional activity of regions to which they are targeted. We performed reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) to identify differential gene expression between modified and control cells. RT-qPCR revealed consistent alteration of TYK2 expression upon inhibition and activation of rs892085. This supports previous observations, where this regulatory region harbouring GWAS-implicated variation operates through direct long-distance contact. To associate genes with disease, the closest or most biological relevant gene is often assigned to a GWAS variant. We have shown that this is not always reliable, with DNA regions looping over large distances to control genes. This provides the first functional validation of the psoriasis-associated SNP rs892085. GWAS have linked TYK2 to psoriasis susceptibility and TYK2 is a recognized drug target with one therapy, deucravacitinib, already approved by the US Food and Drugs Administration. This work demonstrates how risk alleles may work to dysregulate TYK2 signalling in disease. Ultimately, genetic risk scores may be generated to predict how a patient may respond to tyrosine kinase 2 (TYK2) therapy. Functional annotation of loci that perturb the TYK2 pathway will be the key to optimizing TYK2 therapy for the right person at the right time.
O16 The novel roles of laminin α3 in epidermal barrier function and metabolism
Authors: Parzymies Piotr , Jones Eleri Mai , Mayes Andrew  , Messenger David , O’Toole Edel , Caley Matthew 
Published: July 2023

The skin serves an important barrier function performed by epidermal lipids, cornified epithelial cells and cell–cell adhesions. This function is compromised in old age and exhibits certain abnormalities such as flattened dermal–epidermal junction or improper lipid metabolism, and is also associated with mitochondrial dysfunction. The levels of a basement membrane protein laminin 332 have been shown to decrease as the skin ages. We have previously demonstrated that loss of laminin 332 leads to an upregulation of genes involved in cholesterol biosynthesis and is associated with an altered skin lipid profile. We showed that this change is caused by an abnormal intracellular cholesterol transport that is actin-dependent. To further elucidate the mechanism around cholesterol transport, we have identified five proteins of interest for further investigation. We have shown that two of those proteins, NPC2 and Myo5b, are decreased with the loss of the α3 laminin 332 subunit in laminin α3 knockdown nTERT keratinocytes vs. shC controls. Interestingly, the NPC2 binding partner, NPC1, appears to be increased in the laminin α3 knockdown nTERT keratinocytes. Our analysis has also demonstrated that e-cadherin, a crucial cell–cell adhesion protein, as well as several differentiation markers, are altered with the loss of laminin α3. We have carried out laminin 332 recovery experiments in two (2D) and three dimensions, which showed that the addition of recombinant laminin 332 restores the proper levels of myo5b and e-cadherin, suggesting a potential therapeutic effect. We have also performed proteomic, lipidomic, and transcriptomic studies in 2D, which demonstrated statistically significant changes (Q-value ≤ 0.01) in proteins involved in actin and microtubule regulation, cell–cell adhesion and showed unexpected mitochondrial metabolic changes associated with the loss of laminin α3. These findings show that laminin α3 plays numerous, previously unreported roles in epidermal lipid transport, cell–cell adhesion, differentiation and metabolism, and suggests a potential therapeutic role of the protein in skin ageing.
O19 CYLD cutaneous syndrome tumours demonstrate increased NF-κB signalling and diminished collagen organisation.
Authors: Inns Joseph , Rajan Neil
Published: July 2023

CYLD is a tumour-suppressor gene encoding a deubiquitinase, targeting K63- and M1-linked ubiquitin chains and thereby influencing a variety of cellular processes. Loss of function in patients with CYLD cutaneous syndrome (CCS) results in the formation of benign cutaneous cylindromas, spiradenomas and trichoepitheliomas. These CCS skin tumours originate from the hair follicle and appear from adolescence onwards. We used an unbiased proteomics approach to analyse CCS tumour and control samples (n = 8). Label-free quantification of 627 proteins revealed significant changes in 139 proteins with a fold change > 1.75 and P-value <0.05, between control and tumour samples. Gene Ontology analysis identified upregulation of nuclear factor-κB (NF-κB) and immune signalling proteins alongside an increased prevalence of spliceosome proteins in tumour samples. Collagen 7, collagen 4, agrin and thrombospondin-1 were increased in tumour samples, providing, for the first time, an unbiased set of extracellular matrix (ECM) proteins generated by CCS tumour cells. Conversely, CCS tumour samples demonstrated a decreased abundance of proteins associated with collagen organization and fibril formation compared to normal skin. Further analysis of CCS tumours through immunoblotting revealed a reduction of collagen I, while collagen VII was highly upregulated in both tumour lysate and secretome. Our findings indicate that loss of CYLD function impacts on cellular signalling pathways such as NF-κB signalling, and is associated with hypersecretion of select ECM proteins.
O21 Single-cell RNA sequencing and proteomic analysis of dermal fibroblasts cultured from female scalp identifies significant alterations in cell populations with age
Authors: Baker Richard , Williams Rachael , Westgate Gillian  , Pawlus Alison , Zguris Jeanna , Thornton Julie
Published: July 2023

We have previously reported that significant in situ detrimental changes occur in the female scalp with age. We also reported that primary cultures of female scalp dermal fibroblasts (DF) significantly increase the secretion of proteins such as cartilage oligomeric matrix protein/thrombospondin-5 (COMP/TSP5), secreted protein acidic and cysteine rich (SPARC) and smooth muscle actin (ACTA2). This highlights changes in DF cellular identity with age, altering the immediate hair follicle environment, presumably driving hair ageing. To gain a better understanding of age-related molecular and functional changes in DFs, we compared both the transcriptional and proteomic profile of primary cultures of scalp DFs (passage < 3) from female donors (n = 8) aged 21–77 years. Transcriptional profiles were analysed via 10× Genomics scRNAseq. Data profiles were generated by evaluation of cellular lysates by liquid chromatography mass spectrometry proteomics and compared by MS1 precursor intensities using PEAKS software. A total of 16 166 DF cells from young (n = 3) and older (n = 3) donors were sequenced. Across the two populations, 11 transcriptionally distinct clusters were detected, highlighting the heterogeneity in DFs. The percentage of cells in these clusters changed with age, and gene set enrichment analysis ascertained a significant increase of cells in cluster 2, which represented a higher number of gene sets related to fibrosis, senescence, collagen synthesis and degradation. Proteomic analysis of DF cellular proteins (four donors in each age group) identified 11 significantly differentially expressed proteins between the two age groups. Two proteins (serine/threonine-protein kinase 38 and rho GTPase activating protein 21) were significantly reduced in the older cohort, while nine proteins (smoothelin, retinol dehydrogenase 5 tetranectin, oestradiol 17-beta-dehydrogenase 11, CD14, carboxypeptidase M, gamma-enolase, plexin-B1 and endoplasmic reticulum membrane protein complex subunit 4) were significantly elevated in the older cohort. Gene Ontology and STRING analysis of the data revealed modifications to key pathways with age (e.g. collagen and elastic fibre formation, autophagy, ubiquitination and metabolic markers). This study highlights the loss of scalp DF cell signature and identity with age. We hypothesize that these changes impact on the ability of the scalp dermal environment to support and maintain healthy hair follicles, contributing to the age-related changes observed in female hair growth.
P06 Functional MDM2 ablation in HK1.ras/fos-Δ5PTEN transgenic mice results in an elevated, p53-mediated block of carcinogenesis that depends upon p53 expression
Authors: Moula Sofia , Turnbull Gavin , Greenhalgh David 
Published: July 2023

MDM2 is a ubiquitin ligase that targets p53 for degradation and the efficacy of functional MDM2 knockout to protect p53 levels was shown to inhibit benign papilloma formation in HK1.ras transgenic mice. In skin carcinogenesis driven by activated rasHa/fos expression (HK1.ras/fos) and ablation of phosphatase and tensin homolog (PTEN)-mediated Akt regulation, p53 loss was a consistent theme leading to well-differentiated squamous cell carcinoma (SCC). Analysis of endogenous MDM2 status in HK1.ras/fos-Δ5PTENflx hyperplasia and papillomas showed that activated p-MDM2166 activity was initially confined to suprabasal layers with resultant elevated basal layer p53. However, with time, HK1.ras/fos-Δ5PTENflx papillomas exhibited reduced p53 levels in basal keratinocytes associated with increased p-MDM2166 activity, which coincided with malignant conversion, suggesting that suprabasal-to-basal increases in p-MDM2166 downregulated p53 giving a susceptibility to malignant conversion. Therefore, to assess whether removal of this MDM2 ability to degrade p53 could inhibit carcinogenesis in HK1.ras/fos-Δ5PTENflx mice, MDM2 exons 5 and 6, which bind p53 and chaperone for ubiquitination, were removed via a Cre/lox-P system driven by a keratin K14 promoter following treatment with topical RU486 (K14cre-Δ5/6MDM2flx/flx). RU486 treatment of HK1.ras/fos-Δ5PTENflx-Δ5/6MDM2flx/flx mouse skin resulted in a complete block of carcinogenesis in all animals for over 16 weeks due to elevated p53 expression. However, preneoplastic hyperplasia appeared to be relatively unaffected, despite excess p53 expression. In approximately 10–15% of animals, HK1.ras/fos-Δ5PTENflx-Δ5/6MDM2flx/flx mice developed overt tumours by 5–6 months. A similar finding was observed in HK1.fos-Δ5PTENflx keratoacanthoma (KA) aetiology, as here the early p53 expression in HK1.fos-Δ5PTENflx-Δ5/6MDM2flx/flx hyperplasia mediated by MDM2 loss also blocked KA aetiology. To assess whether this was specific to MDM2/p53 interactions, inducible p53 ablation was introduced into the HK1.fos-Δ5PTENflx KA model. This confirmed inhibition was specific to p53 expression, as all HK1.fos-Δ5PTENflx/p53 flx/flx tumours rapidly progressed to malignancy, regardless of MDM2 genetic status. Thus, MDM2 inhibition can limit carcinogenesis via maintaining p53 levels; however, this inhibition can be circumvented via p53 loss and, moreover, MDM2 inhibition would be counterproductive in SSC contexts that expressed gain-of-function p53 mutations.
P26 Characterization of skin barrier defects in atopic dermatitis patients using attenuated total reflection Fourier transform infrared spectroscopy
Authors: Williams Samuel , Danby Simon
Published: July 2023

Atopic dermatitis (AD) is a common disorder characterized by skin barrier defects often measured by traditional biophysical tools that give little molecular insight into disease pathogenesis. The objective was to employ in vivo infrared spectroscopy as a complimentary technique to biophysical measurements to analyse changes in the chemical composition of the stratum corneum (SC) in relation to AD severity. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy measurements were collected on the forearm alongside transepidermal water loss (TEWL), skin surface pH and capacitance in a cohort of 75 participants, stratified by AD severity. Skin tape stripping was performed to allow measurements at greater SC depth. The five common filaggrin (FLG) loss-of-function mutations were genotyped. Skin hydration, TEWL and redness were all affected by AD severity with molecular analysis showing concomitant changes in lipid structure, and the amount of lipid, polyamide and carboxylate. Correlation analysis showed associations between Eczema Area and Severity Index and lipid structure (r = –0.58), as well as amount of carboxylate (r = –0.49). Principal component analysis showed spectra collected on participants with AD lesions or FLG mutations clustered away from healthy and nonlesional skin sites. Our findings showed that ATR-FTIR spectroscopy is able to characterize structural skin barrier defects in people with AD vs. controls. Spectral peak analysis showed associations between defective lipid and natural moisturizing factor (NMF) components with worsening disease severity. Through combined assessment of peaks associated with lipid structure and carboxylate amount, we have shown the potential of ATR-FTIR in identifying filaggrin deficiency, through either genetic predisposition (FLG mutations) or through inflammatory action, which is related to the presence of NMFs in the SC, skin hydration and overall severity of AD.
P31 Dissecting the epigenetic and transcriptional regulators in keloid scars
Authors: Tekkela Stavroula , Drudi Elena M , Shaw Tanya , Philpott Mike , O’Toole Edel , Rognoni Emanuel
Published: July 2023

Keloids are a type of fibroproliferative scar that progressively grow beyond the original wound site. Currently, there are no treatments to halt or revert keloid scarring, emphasizing the great unmet clinical need and incomplete mechanistic understanding. They occur due to genetic, epigenetic and environmental abnormalities. However, the detailed molecular signatures that promote this pathogenic phenotype are still unknown. Fibroblasts are best characterized for their role in depositing and remodelling the extracellular matrix and presumed to be one of the main cells involved in keloid. This study investigates if keloid fibroblasts are epigenetically locked in an activated state, and if manipulation of this state can reverse the persistent keloid phenotype. To investigate the epigenetic and genetic changes in keloid fibroblasts, we performed ATACseq and RNAseq on a patient-matched set of keloid and normal fibroblasts. ATACseq is a high-throughput sequencing method that defines chromatin-accessible regions in which transcription factor binding sites can be mapped that can regulate gene expression. Differences in chromatin accessibility were found to be closely associated with gene expression differences in keloid fibroblasts. ATACseq analysis revealed accessible regions at genes that are implicated in pathways, including transcriptional regulation, embryonic development, calcium binding, collagen remodelling and apoptosis. Identified candidates driving keloid formation will be validated in keloid skin sections and functionally analysed in two- and three-dimensional culture models after pharmacological or genetic manipulation. These findings will support the development of fibroblast-targeted therapies for keloid scars and possibly other connective tissue disorders.
Time-based reperfusion in haemodynamically unstable pulmonary embolism patients: does early reperfusion therapy improve survival?
Authors: Zuin Marco , Piazza Gregory , Barco Stefano , Bikdeli Behnood , Hobohm Lukas , Giannakoulas George , Konstantinides Stavros
Published: July 2023

High-risk pulmonary embolism (PE) is associated with significant morbidity and mortality. Systemic thrombolysis remains the most evidenced-based treatment for haemodynamically unstable PE, but in daily clinical practice, it remains largely underused. In addition, unlike acute myocardial infarction or stroke, a clear time window for reperfusion therapy, including fibrinolysis, for high-risk PE has not been defined either for fibrinolysis or for the more recently incorporated options of catheter-based thrombolysis or thrombectomy. The aim of the present article is to review the current evidence supporting the potential benefit of earlier administration of reperfusion in haemodynamically unstable PE patients and suggest some potential strategies to further explore this issue.