Network Analysis Research Articles

Comprehensive bioinformatic analysis identified m6A methylation as an important regulator in mediating immune response during idiopathic pulmonary arterial hypertension

Abstract Background Despite its functional importance in various fundamental bioprocesses, studies of N6-methyladenosine (m6A) in the pulmonary arterial hypertension (PAH) are lacking. Here we studied the potential relevance of m6A RNA methylation and immune response in PAH development. Methods We constructed a monocrotaline (MCT) induced PAH rat model and performed Methylated RNA immunoprecipitation sequencing (MeRIP-Seq). The 18 idiopathic PAH (IPAH) microarray data obtained from the GEO database was used to construct co-expression networks by weighted gene co-expression network analysis (WGCNA). CIBERSORT was used to investigate the effect of m6A methylation on immune cell infiltration during PAH. Results A differential pattern of m6A abundance, mainly up-methylation, was observed in the lung tissues of rats with MCT induced PAH. By WGCNA, multi-list pathway enrichment analysis and protein-protein interaction (PPI) analysis, we found that m6A methylation modification may play important roles in mediating immune response during PAH. CYBERSORT algorithm indicated that the m6A methylation can drive monocyte to form M1 macrophage, which may be mediated by CCR5 and CXCL9. Conclusion Collectively, m6A landscape is altered in PAH. We summarize newly discovered m6A in controlling immune response, which caused activation of M1 macrophage during PAH. It’s provided a novel insight into the therapeutic mechanisms of PAH.

Tobacco Smoking Functional Networks: A Whole-Brain Connectome Analysis in 24,539 Individuals.

Nicotine addiction, a multifaceted neuropsychiatric disorder, profoundly impacts brain functions through interactions with neural pathways. Despite its significance, the impact of tobacco smoking on the whole-brain functional connectome remains largely unexplored. We conducted a whole-brain analysis on 24,539 adults aged 40 and above from the UK Biobank cohort. Subjects were categorized into individuals who use nicotine and who do not use nicotine based on current and chronic tobacco smoking information. Functional connectivity was assessed using resting-state functional magnetic resonance imaging (rfMRI). We employed a network analysis method to assess the systematic effects of tobacco smoking on brain connectome by identifying subnetworks that show nicotine-use-related differences. Our analyses revealed two nicotine-use-related subnetworks with distinct network structure (permutation p-value < 0.001). In the first network, there is a significant decrease in resting-state functional connectivity (rsFC) between the basal ganglia regions (e.g., nucleus accumbens) and 73% of the remaining brain regions, emphasizing the central hub role of basal ganglia in addictive smoking behaviors. Additionally, a data-driven subnetwork mainly involving regions from frontal and occipital lobes, showed reduced rsFC among individuals who use nicotine. The results suggest significant alterations in the communication and coordination among the basal ganglia and the broader network of brain regions. The observed changes in rsFC indicate a widespread disruption in the connectivity patterns associated with nicotine use. This study identifies rsFC subnetworks related to chronic nicotine use through whole-brain connectome analysis. The findings confirm that widespread alterations in rsFC are centered around hub nodes within the basal ganglia, including bilateral nucleus accumbens, putamen, caudate, and globus pallidus. In addition, our analysis found a clique-forming subnetwork vulnerable to tobacco smoking consisting of regions from the visual, dorsal/ventral attention, and frontoparietal networks.

Mechanistic exploration of bioactive constituents in Gnetum gnemon for GPCR-related cancer treatment through network pharmacology and molecular docking

G Protein-Coupled Receptors (GPCRs) are integral membrane proteins that have gained considerable attention as drug targets, particularly in cancer treatment. In this study, we explored the capacity of bioactive compounds derived from Gnetum gnemon (GG) for the development of of pharmaceuticals targeting GPCRs within the context of cancer therapy. Integrated approach combined network pharmacology and molecular docking to identify and validate the underlying pharmacological mechanisms. We retrieved targets for GG-derived compounds and GPCRs-related cancer from databases. Subsequently, we established a protein-protein interaction (PPI) network by mapping the shared targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were employed to predict the mechanism of action of these targets. Molecular docking was conducted to validate our findings. We identified a total of 265 targets associated with GG-derived bioactive compounds for the treatment of GPCRs-related cancer. Functional enrichment analysis revealed the promising therapeutic effects of these targets on GPCRs-related cancer pathways. The PPI network analysis identified hub targets, including MAPK3, SRC, EGFR, STAT3, ESR1, MTOR, CCND1, and PPARG, which demonstrate as treatment targets for GPCRs-related cancer using GG-derived compounds. Additionally, molecular docking experiments demonstrated the strong binding affinity of gnetin A, gnetin C, (-)-viniferin, and resveratrol dimer, thus inhibiting MAPK3, SRC, EGFR, and MTOR. Survival analysis established the clinical prognostic relevance of identified hub genes in cancer. This study presents a novel approach for comprehending the therapeutic mechanisms of GG-derived active compounds and thereby paving the way for their prospective clinical applications in the field of cancer treatment.

Experimental and in silico analysis of LINC01279 expression in tumor of patients with breast cancer.

Breast cancer (BC) is characterized by the increase of malignant cells in the breast. The malignant cells begin in the lining of the breast milk glands or ducts (ductal epithelium). BC is the most frequent cancer in women, but it may also occur in males. Long non-coding RNAs (lncRNA) have been demonstrated to control the development and incidence of cancer. However, some lncRNAs experience potential changes in BC, but their role has not been well studied. LINC01279 is known as a valuable biomarker in gastric cancer but has not yet been studied in BC. Changes in LINC01279 expression levels in BC samples were investigated by microarray. Q-PCR was also used to evaluate the expression of LINC01279 in the tumor and normal adjacent samples of 30 BC patients. The LINC01279 co-expressed gene module was discovered using weighted gene correlation network analysis (WGCNA) on the relevant dataset. The top ten hub genes were determined using gene ontology (GO) functional enrichments on the co-expressed gene module. The results of the bioinformatics study showed an increase in LINC01279 expression levels (log2FC = 3.228749561, adj.P.Val = 1.69E - 12) in tumor samples compared to normal marginal tissue. Q-PCR results also showed a significant increase in LINC01279 expression (P-value = 0.0005) in tumor samples. WGCNA analysis identified that the black module is the LINC01279 co-expressed module, and functional annotation analysis of black module genes enriched in significant cancer-related pathways and processes, including cell growth and/or maintenance, regulation of immune response, regulation of cell proliferation, and epithelial-to-mesenchymal transition (EMT). Regarding the real-time PCR results, the analysis of expression patterns has illuminated a distinct association between the heightened expression levels of LINC01279, and the stages of cancer progression as well as the metastatic potential of tumors. However, intriguingly, our observations have failed to reveal any statistically significant correlations between the relative expression of LINC01279 and tumor grade classification, or the presence of ER, PR, and HER2 biomarkers. The present study could provide a new perspective on the molecular regulatory. Processes associated with BC pathogenic mechanisms are linked to the LINC01279, although further research is needed on the possible role of this lncRNA in BC.

Combined Transcriptome and Metabolome Analysis Reveals That Carbon Catabolite Repression Governs Growth and Pathogenicity in Verticillium dahliae

Carbon catabolite repression (CCR) is a common transcriptional regulatory mechanism that microorganisms use to efficiently utilize carbon nutrients, which is critical for the fitness of microorganisms and for pathogenic species to cause infection. Here, we characterized two CCR genes, VdCreA and VdCreC, in Verticillium dahliae that cause cotton Verticillium wilt disease. The VdCreA and VdCreC knockout mutants displayed slow growth with decreased conidiation and microsclerotium production and reduced virulence to cotton, suggesting that VdCreA and VdCreC are involved in growth and pathogenicity in V. dahliae. We further generated 36 highly reliable and stable ΔVdCreA and ΔVdCreC libraries to comprehensively explore the dynamic expression of genes and metabolites when grown under different carbon sources and CCR conditions. Based on the weighted gene co-expression network analysis (WGCNA) and correlation networks, VdCreA is co-expressed with a multitude of downregulated genes. These gene networks span multiple functional pathways, among which seven genes, including PYCR (pyrroline-5-carboxylate reductase), are potential target genes of VdCreA. Different carbon source conditions triggered entirely distinct gene regulatory networks, yet they exhibited similar changes in metabolic pathways. Six genes, including 6-phosphogluconolactonase and 2-ODGH (2-oxoglutarate dehydrogenase E1), may serve as hub genes in this process. Both VdCreA and VdCreC could comprehensively influence the expression of plant cell wall-degrading enzyme (PCWDE) genes, suggesting that they have a role in pathogenicity in V. dahliae. The integrated expression profiles of the genes and metabolites involved in the glycolysis/gluconeogenesis and pentose phosphate pathways showed that the two major sugar metabolism-related pathways were completely changed, and GADP (glyceraldehyde-3-phosphate) may be a pivotal factor for CCR under different carbon sources. All these results provide a more comprehensive perspective for further analyzing the role of Cre in CCR.

Novel metabolomic predictors of incident colorectal cancer in men and women.

Metabolomic profiles may influence colorectal cancer (CRC) development. Few studies have performed pre-diagnostic metabolome-wide analyses with CRC risk. We conducted a nested case-control study among women (Nurses' Health Study (NHS)) and men (Health Professionals Follow-up Study (HPFS)) who provided blood between 1989 and 1995. Over 22.9 years, 684 (409 NHS, 275 HPFS) incident CRC cases occurred and were matched 1:1 to controls. Liquid chromatography-mass spectrometry (LC-MS) identified 255 plasma metabolites after quality control. Cohort-specific and combined metabolite association analyses were performed using conditional logistic regression. Metabolite set enrichment analysis (MSEA) was used to identify differential abundance in metabolite classes. Weighted Correlation Network Analysis (WGCNA) provided modules of covarying metabolites, which were tested for CRC association. MSEA identified specific acylcarnitines associated with higher CRC risk and triacylglycerols with lower CRC risk among women and men. Further, phosphatidylcholines were associated with a higher risk of CRC among men. In an analysis restricted to CRC cases diagnosed two years after blood draw, myristoleic acid (OR = 1.37; 95%CI = 1.15-1.62; FDR = 0.072) and C60:12 triacylglycerol (OR = 0.75; 95%CI = 0.64-0.88; FDR = 0.072 were associated with CRC risk in women. WGCNA identified amino acids associated with CRC in men, fatty acid esters (carnitines) with distal CRC in men, and triradylcglycerols inversely associated with CRC in women. We identified pre-diagnostic CRC-associated metabolites with distinct sex-specific profiles. These results provide insight into CRC etiopathogenesis and have implications for risk prediction strategies.

Multi-omics integration analysis identifies pathogenic candidate genes of congenital heart defects with vascular malformation

Abstract Background Congenital heart defects (CHD) with vascular malformation harbors numerous genetic and epigenetic aberrations of DNA copy numbers and methylation. Proteomic deregulation by the aberrations plays key driver roles in heterogeneous progression of CHD. However, it still remains largely unknown about integrative analysis of genetic, epigenetic regulation and proteome in CHD. Purpose We employed a multi-omics approach to explore pathogenic candidate genes in CHD with vascular malformation. Methods Previously, we gathered WES data for 216 CHD patients with vascular malformation and 100 healthy controls. Meanwhile, we carried out DNA methylation analysis from 11 patients and 5 controls. Here, we profile proteomics of vascular tissues from 11 patients and 5 controls. Integration analysis of DNA copy-number-correlated (CNVcor) genes, methylation-correlated (METcor) genes and differentially proteins reveal the innovative and crucial pathogenetic genes. We then detected spatiotemporal expression patterns of candidate genes in zebrafish embryos using in situ hybridization and qPCR, and verified the roles in vascular development by morpholino-mediated gene knockdown. Results 90 differentially expressed proteins (DEPs) were obtained using proteomics and were mainly related to cell adhesion and migration, angiogenesis, amino acid metabolism, and HIF-1 signaling pathway through GO and KEGG pathway enrichment analysis. 15 hub proteins were identified by WGCNA analysis (Fig 1). Additionally, we found that the frequencies of the aberrancies of the CNVcor genes were significantly correlated with the METcor genes (Fig 2 C). We obtained 1924 differentially CNVcor genes and 952 differentially METcor genes. Then, we computed the Pearson’s correlation coefficients of all three-omics by pairwise comparison and found that the distribution of correlations was bimodal (Fig 2 D). Meanwhile, 28 overlapped genes were observed among three-omics. We screened out 11 candidate genes (TIMP1, BGN, LTBP1, AKR1C3, LOXL3, MAOA, MAOB, GNA12, IFI30, CNTNAP1, IGF2BP1) according to the node degree of PPI network analysis between 28 genes and known vascular development related genes. We further figured out LOXL3, MAOA, CNTNAP1 and IGF2BP1 are the novel important genes for vascular malformation, of which CNTNAP1 is the joint gene of three-omics. Through expression patterns and gene knockdown in zebrafish embryos, we revealed that LOXL3, MAOA, CNTNAP1 and IGF2BP1 affect vascular development (Fig 2 E-G). Conclusions We identify novel key candidate genes (LOXL3, MAOA, CNTNAP1 and IGF2BP1) potentially related to CHD with vascular malformation and elucidate the possible molecular pathogenesis of CHD. Our multi-omics integrative analysis provides new insights on the multi-layered etiology of CHD, which might be helpful in developing clinical diagnosis and precaution for CHD.Figure 1Figure 2

Identification of genes associated with sex expression and sex determination in hemp (Cannabis sativa L.).

Dioecy in flowering plants has evolved independently many different times, and thus the genetic mechanisms underlying sex determination are diverse. In hemp (Cannabis sativa), sex is controlled by a pair of sex chromosomes (XX for females and XY for males). In an attempt to understand the molecular mechanism responsible for sex expression in hemp plants, we carried out RNA-Seq of male and female plants at different developmental stages. Using a pipeline involving differential gene expression analysis and weighted gene co-expression network analysis, we identified genes important for male and female flower development. We also demonstrate that sex-biased expression is already established at very early vegetative stages, before the onset of reproductive development, and several genes encoding transcription factors of the REM, bZIP and MADS family as candidate sex determination genes in hemp. Our findings demonstrate that the gene regulatory networks governing male and female development in hemp diverge already at a very early stage, leading to profound morphological differences in male and female hemp plants.

Using Social Network Analysis during COVID-19 contact tracing in Libya, 2020

Abstract Background Social network analysis (SNA) may provide valuable insights into individual-level transmission dynamics, allowing for targeted contact tracing interventions. We used SNA to understand the variability in SARS-CoV-2 transmission at the case level during the first wave in Libya. Methods We analyzed the spread of COVID-19 in Libya from March to August 2020 using contact tracing data from 287 confirmed cases. We created a network to visualize the transmission patterns, representing each case as a node and connections as links. We focused on direct contacts identified through contact tracing and calculated network measures such as out-degree centrality, in-degree centrality, and betweenness centrality. Gephi software was used for data visualization and understanding the network structure. Results Of the 287 COVID-19 cases, 264 (92%) had nodes with zero outdegrees, indicating that they did not contribute to infection transmission. The remaining 23 (8%) had non-zero out-degree measures and transmitter infection to other cases, 10 (4%) of whom were super-spreaders (outdegree≥5) causing 70% of the transmission. Six (2%) had no epidemiological contact, indicated by zero in-degree centrality measures; 63% had zero betweenness centrality measures, suggesting they did not act as intermediaries connecting separate transmission chains within the network. The network contained nine clusters of connected nodes, with the largest having 91 (24%) cases; the mean path length between cases was 1.846, and the network diameter was 4. Conclusions The study revealed distinct clusters of connected cases, along with the presence of super-spreaders. This network structure underscores the critical role of early identification and isolation of individuals with high transmission potential to effectively contain outbreaks. This revised conclusion directly references the findings of nine clusters and super-spreaders, demonstrating how these results support the importance of early intervention. Key messages • Social network analysis identifies clusters and super-spreaders in COVID-19 transmission, emphasizing the need for targeted interventions. • Connected clusters and super-spreaders highlight the importance of contact tracing and early isolation for effective control of COVID-19 outbreaks.

Transcriptomic analysis of cellular senescence induced by ectopic expression of ATF6α in human breast cancer cells.

The transcriptomic profile of cellular senescence is strongly associated with distinct cell types, the specific stressors triggering senescence, and temporal progression through senescence stages. This implies the potential necessity of conducting separate investigations for each cell type and a stressor inducing senescence. To elucidate the molecular mechanism that drives endoplasmic reticulum (ER) stress-induced cellular senescence in MCF-7 breast cancer cells, with a particular emphasis on the ATF6α branch of the unfolded protein response. We conducted transcriptomic analysis on MCF-7 cells by ectopic expression of ATF6α. Transcriptomic sequencing was conducted on MCF-7 cells at 6 and 9 hours post senescence induction through ATF6α ectopic expression. Comprehensive analyses encompassing enriched functional annotation, canonical pathway analysis, gene network analysis, upstream regulator analysis and gene set enrichment analysis were performed on Differentially Expressed Genes (DEGs) at 6 and 9 hours as well as time-related DEGs. Regulators and their targets identified from the upstream regulator analysis were validated through RNA interference, and their impact on cellular senescence was assessed by senescence-associated β-galactosidase staining. ATF6α ectopic expression resulted in the identification of 12 and 79 DEGs at 6 and 9 hours, respectively, employing criteria of a false discovery rate < 0.05 and a lower fold change (FC) cutoff |log2FC| > 1. Various analyses highlighted the involvement of the UPR and/or ER Stress Pathway. Upstream regulator analysis of 9 hour-DEGs identified six regulators and eleven target genes associated with processes related to cytostasis and 'cell viability and cell death of connective tissue cells.' Validation confirmed the significance of MAP2K1/2, GPAT4, and PDGF-BB among the regulators and DDIT3, PPP1R15A, and IL6 among the targets. Transcriptomic analyses and validation reveal the importance of the MAP2K1/2/GPAT4-DDIT3 pathway in driving cellular senescence following ATF6α ectopic expression in MCF-7 cells. This study contributes to our understanding of the initial molecular events underlying ER stress-induced cellular senescence in breast cancer cells, providing a foundation for exploring cell type- and stressor-specific responses in cellular senescence induction.

8.A. Scientific session: Engage, co-create, collaborate, transform: uniting practice and research for health innovation

Abstract Background Public health practitioners are increasingly challenged by theoretical, political, and contextual changes and innovations. To ensure that innovations are sustainable, it is important not only to include stakeholders (e.g. end-users, implementers, policy-makers), but also to actively involve them in the development, implementation and transformation of public health innovations. Participatory action research, co-creation and stakeholder engagement are proven methods to co-produce effective innovations, strengthen existing networks and promote implementation by increasing fit and applicability in practice. In this workshop, our overall aim is to bring together practitioners and researchers to improve mutual understanding and adapt practices in development, implementation and transformation of innovations. The objectives of the workshop are: 1) to increase participants’ understanding of the circumstances under which stakeholder involvement is most meaningful, and 2) to increase participants’ awareness of tools, strategies or theories to overcome challenges in stakeholder involvement. Workshop format The workshop consists of two parts: four interactive presentations that introduce important development steps of public health innovation and a problem-based discussion using a real-life case that builds upon the presentations. Interactive presentations describe (1) Theory of Change (ToC) approach for active collaboration with stakeholders, (2) co-creation of a serious game for and with healthcare providers, (3) visualising collaboration through Social Network Analysis to improve implementation, and (4) enhancing scale up with PRACTIS and Design Thinking. During the presentations, interactive tools are used to help identify the characteristics of the workshop participants (e.g. work setting, type of stakeholders), as well as their interests and needs (e.g. challenges faced, methods used). During the second part of the workshop, a health innovation is chosen from a number of pre-selected innovations from European contexts (e.g. BOB campaign, Schools4Health, LETHE) that aligns most with participants’ background and interests. Next, the main goal and known challenges within these innovations are introduced, which aids participants to formulate intermediate goals using the ToC approach. Then, they answer questions and make decisions for the discussed stages of innovation development (e.g. “Which gaps exist in the current network and how can we address them to improve collaboration?”, ‘What scalability challenges exist in this innovation and how can PRACTIS help identify them?”). In answering these questions, participants will share their knowledge and expertise with each other, while the chair facilitates the discussion. We are flexible in the format of the challenge-based discussion; depending on the number of attendees, we may work with a plenary discussion or smaller groups that work independently under the supervision of 1 or 2 facilitators each. Key messages • Non-linear approaches are needed to take into account the complicated and changing systems in which health innovation takes place. • Theory of change, co-creation, social network analysis, PRACTIS and design thinking offer practical guidance for stakeholder involvement in specific phases of health innovations.

Relationship network of safety management elements in the construction industry under the perspective of resilience

PurposeConstruction safety resilience is gradually gaining attention in the field of engineering construction as a new management concept and way to improve safety performance. However, how to cope with the dilemma of the unclear relationship of construction safety resilience elements at the practice level and promote the harmonization of construction safety goals and resilience enhancement paths has become an urgent challenge for safe construction.Design/methodology/approachThis study analyzes the components of construction safety resilience elements. A relationship network model of construction safety resilience elements is developed by using the social network analysis method. The location and influence of each element in the network and the interrelationships among the elements are explored in depth.FindingsThe findings reveal a robust interconnection among the elements of safety resilience in the construction industry. Key components such as safety behavior, risk prevention and control mechanisms, disaster prevention and mitigation technologies as well as information technology, are positioned at the core of the network. Notably, safety behavior exerts the most significant influence over the other elements, serving as the linchpin of safety management in the construction industry. Moreover, the interplay among safety resilience elements in the construction sector can alter the structure of the relationship network.Originality/valueThis study adopts the social network approach to solve the problem that it is difficult to quantitatively analyze the elements of construction safety resilience and their interrelationships and to clarify the interactions among the core elements, which can help to further assist the construction project manager to continuously optimize safety resilience and improve construction safety.

Preparedness for and collaboration during regional outbreaks with multidrug-resistant organisms

Abstract Background Outbreaks with multidrug-resistant organisms (MDROs) are a significant burden on the healthcare system and the wellbeing of affected patients and carriers. Cross-institutional MDRO outbreaks require a concerted multidisciplinary effort for effective outbreak management. This presents challenges regarding cooperation, coordination and communication between stakeholders. In this study, we investigate collaborative structures and coordination roles in Dutch outbreak response networks and how Dutch healthcare professionals engage in preparedness activities relating to MDRO outbreaks. These findings form the basis for the development of a stress test exercise for improving professional and organizational collaboration and coordination during MDRO outbreaks. Methods Using a mixed-methods design, an online survey and qualitative interviews are conducted among healthcare professionals in two healthcare regions in the Netherlands. Social network analysis is used to gain insight in collaborative structures and brokerage roles. To investigate preparedness activities, the survey is combined with qualitative interviews to explore respondents’ experiences, preferences and needs regarding preparedness activities for MDRO outbreak management. Results During the workshop we will show the cross-organizational network structures, the strengths and weakness of the mutual connections and provide insight in positions in networks, roles and responsibilities. Also, we provide insight in how the use of and experience with training and exercise activities, such as stress tests, contribute to preparedness for MDRO outbreaks in vulnerable patient populations. Conclusions This study shows how healthcare professionals from different organizations collaborate during MDRO outbreaks, and how they engage with preparedness activities. This provides valuable input for development of a stress test specific to MDRO outbreak management, and other interventions to improve outbreak preparedness.

Development of a Large-Scale Horizontal Curve Inventory for Safety Analysis Using Open GIS Data

Abstract Horizontal curves are crucial for the safety and mobility of roadways. To enhance the understanding of these curves, transportation agencies are investigating methods to accurately determine their spatial locations, geometric attributes, and conditions within their extensive road networks. Leveraging the widespread availability of Global Positioning System (GPS) and Geographic Information System (GIS) data, significant advancements have been made in developing automated algorithms that identify horizontal curves from GPS trajectories and GIS basemaps. This study introduces a novel dataset—the Horizontal Curve Inventory—that encapsulates detailed features of horizontal curves. We employed a modified automated cycle regression algorithm designed for large-scale road network analysis to extract horizontal curve data. This method was applied nationally to process an expansive network utilizing open-source GIS basemaps, encompassing over 300,000 miles across interstate, U.S. highway, and state highway systems. The inventory includes comprehensive data on curve radius, rotation angle, start and end point coordinates, and the curve’s center points. These data are essential for supporting safety research pertaining to road alignment. Our approach demonstrated robust performance across various open GIS data sources and provides the first accurate, efficient, and comprehensive inventory of horizontal curves for all major highway systems in the United States. The dataset is available at http://aichengbo.com/research/national-horizontal-curve-inventory/.

From mass marketing to personalized digital marketing in tourism: a 2050 horizon paper

Purpose The academic background of tourism marketing dates back to the 1960s. There had been a slight increase in its capacity until the early 1990s. However, since then, it has boomed, reaching thousands of scientific journal articles and tens of scientific books published only in English each year. Therefore, this study aims to present how tourism marketing has progressed academically within the past 60 years over four waves and how this progress may move forward as the next wave. Design/methodology/approach A bibliometric analysis grounds this study, which characterizes the past and present of tourism marketing research and anticipates the future. Content analysis, such as word clouds and social network analysis, was adopted to identify topic clusters and their connections. A total of 9,239 articles published between 1969 and 2024 were extracted from Scopus. Software packages such as VOSviewer were used to determine connections within topics. Findings The authors have assessed the findings broadly. Four waves were from the late 1960s to the first quarter of 2000. In spite of the limited size of papers at the early stage, the last wave saw a boom and a diversified number and category of subjects studied. In each wave, new subjects were added to broaden the picture. Research limitations/implications The discussion of findings is based only on those scientific papers published in English since 1969 but excludes the whole list of textbooks. Subsequent research should also consider all papers and textbooks released in different languages to have the broadest picture assessment worldwide. Practical implications The study conveys various suggestions for industry practitioners and policymakers to focus on carefully assessing trends in marketing tourism services and how these may be shaped shortly. This may help practitioners and policymakers redesign their services and marketing strategies in light of future developments. Originality/value This study continues a recent study published by Kozak (2023) that has been elaborated, particularly from the industry perspective. The current study examines the academic profile of all subjects investigated in the case of tourism marketing, but mainly in an academic sense. Accordingly, this paper outlines the facts and trends researchers may follow throughout the research frame published in the past six decades.

Adherence to Mediterranean diet and cardiometabolic risk among mountain communities in European Alps

Abstract Background The Mediterranean diet has emerged as a model promoting a better health status, particularly for cardiovascular diseases prevention. Adherence to the Mediterranean diet (AMD) may vary depending on geographical location and sociocultural factors. Mountains and rural communities may experience lower adherence, also depending on food provision-related services availability. Our cross-sectional study aims to investigate the association between AMD and cardiometabolic risk in a cohort of mountain dwellers. As a secondary aim, we will test if services availability may influence health status via AMD. Methods In spring/summer 2023, healthy volunteers from 5 small villages in Italian Alps were enrolled. Dietary habits were investigated by Medi-Lite standardized questionnaire. The waist-to-height ratio (WHtR) was calculated by dividing the waist circumference by height, then used as an anthropometric index for central adiposity and related cardiometabolic risk. Services availability was assessed by network analysis implemented in QGIS. A multilevel logistic regression model, with center as random intercept and adjusted for age, sex, BMI, education and physical activity was used to test the association between AMD and cardiometabolic risk. Ethical approval was provided by University of Turin. Results To date, 142 subjects (59.2% females, 49.9±14.0 y.o.) have been enrolled. Preliminary analyses showed that, as compared to the least adherent group, subjects reporting the highest AMD level (3rd tertile of Medi-Lite scoring) were strongly protected from cardiometabolic risk (OR: 0.07, 95% C.I.: 0.01, 0.67; p = 0.021). A similar trend was observed for the 2nd tertile. Conclusions Our preliminary findings suggest the beneficial role of the Mediterranean diet also in remotely located populations. We plan to investigate how service availability may impact AMD and, indirectly, cardiometabolic health to reinforce the importance of equity in services distribution for Public Health Key messages • Independently of age, sex, BMI, education and physical activity, adherence to the Mediterranean diet can promote cardiometabolic health among rural and mountain communities. • Mediterranean eating pattern can positively influence central adiposity and should be promoted to ensure more effective prevention strategies in mountain dwellers.

4.C. Scientific session: Navigating through complex challenges of responding to outbreaks of multidrug-resistant organisms

Abstract Antimicrobial resistance is one of the top ten global public health threats according to the WHO, contributing to millions of deaths each year. While outbreaks of multidrug-resistant organisms (MDROs) are well studied in hospital and nursing home settings, knowledge gaps exist concerning the complex challenges of outbreaks of MDROs among vulnerable patient populations, such as patients with intellectual disabilities or elderly patients living longer at home with multiple comorbidities. The coordination of such outbreaks requires a multidisciplinary approach as these patients often receive care throughout different healthcare settings, including long-term care and home-based nursing care. We delve into the complexities of coordinating an MDRO outbreak response involving healthcare professionals from various healthcare settings. The challenges inherent to the coordination of multidisciplinary involvement and the use of innovative strategies for outbreak preparedness and surveillance, will be thoroughly examined based on scientific evidence and practice based experiences. Workshop objectives • To exchange experiences on managing cross-institutional outbreaks of MDROs • To foster deeper understanding of the complexities of multidisciplinary involvement during MDRO outbreaks among vulnerable patient populations The workshop structure: - A brief introduction will be provided on the generic challenges in providing long-term care to vulnerable patient populations carrying MDROs. We will subsequently illustrate potential challenges faced during a cross-institutional MDRO outbreak among these patients through a realistic scenario (5 min): A patient with severe intellectual and physical disabilities living in both a long-term care facility as well as with family at home, unexpectedly turns out to be carrying an MDRO after recent hospital admission. As the patient receives nursing care from both from family members and nursing staff in different healthcare settings, a cross-institutional MDRO outbreak has occurred requiring a coordinated response. - Hereafter, individual presenters will address evidence- and experience-based challenges faced in MDRO outbreak responses within the specific healthcare settings covered in the previously presented scenario. We will navigate through the scenario together with the participants, discussing practical and ethical dilemmas, and explore ways for containing the MDRO outbreak while maintaining effective and compassionate healthcare delivery. In addition, we will discuss the use of public health innovations, such as social network analysis as a basis for the development of a stress test exercise to improve preparedness for MDRO outbreaks, and the role of wastewater surveillance in MDRO outbreaks. Discussions will be facilitated by (online) voting tools to invite participants to exchange experiences (40 min). We conclude with an exciting quiz to summarize the key messages of this workshop (10 min). Key messages • Healthcare setting, knowledge, proportionality and well-being of vulnerable patients populations are important to acknowledge during cross-institutional MDRO outbreaks. • Novel stress test exercise and wastewater surveillance are potential methods to improve preparedness and surveillance, respectively, during cross-institutional MDRO outbreak responses.

Evaluation on the Efficacy of Farrerol in Inhibiting Shoot Blight of Larch (Neofusicoccum laricinum)

Neofusicoccum laricinum is the causal agent of larch shoot blight, a fungal disease affecting several species of larch. It causes severe damage, including stunting and mortality. This study aims to address the severe impact of larch shoot blight by evaluating the effect of farrerol on the inhibition of Neofusicoccum laricinum in Larix olgensis. We used LC-MS/MS and weighted gene co-expression network analysis to investigate farrerol’s effects on Neofusicoccum laricinum and identify associated genes in resistant and susceptible larch. Our study identified significant differences in metabolite profiles between resistant and susceptible cultivars, with higher concentrations of farrerol showing complete inhibition of N. laricinum. Additionally, specific genes associated with farrerol content were up-regulated in resistant larch. Farrerol at higher concentrations completely inhibited N. laricinum, showing a strong correlation with increased disease resistance. This research suggests that farrerol enhances disease resistance in larch and provides a foundation for developing disease-resistant larch varieties based on antifungal metabolite traits.

Soil bacterial community characteristics and influencing factors in different types of farmland shelterbelts in the Alaer reclamation area

To investigate the effects of various types of farmland shelterbelts on soil quality and soil bacterial community diversity, this study focused on soil samples from four different shelterbelt types in the Alaer reclamation area, including Populus euphratica Oliv.- Populus tomentosa Carrière (PP), Elaeagnus angustifolia L.- Populus euphratica Oliv. (EP), Populus alba var. pyramidalis Bunge (P), and Salix babylonica L. (S). We analyzed their physical, chemical, biological properties as well as the differences in bacterial community structure, and explored the influencing factors on soil microbial community characteristics through microbial correlation network analysis. The results showed that: (1) There were significant differences in soil properties among the four types of farmland shelterbelts (p &amp;lt; 0.05), with P soils exhibiting the highest levels of organic matter, total nitrogen, and total phosphorus contents. (2) The Alpha diversity indices of soil bacteria showed significant differences among the four types of farmland shelterbelts (p &amp;lt; 0.05), with the P soils displayed the highest Chao1 and Shannon indices. (3) There were differences in the composition and abundance of dominant soil bacterial communities among different farmland shelterbelts, notably, the abundances of Verrucomicrobia, Acidobacteria, and Planctomycetes were significantly higher in P soils compared to the other three types. (4) The complexity of the correlation network between microbial species and environmental factors was highest in EP soils, soil microbial biomass nitrogen and available phosphorus were the main influencing factors. These findings indicated that different types of farmland shelterbelts had significant impacts on soil properties and soil bacterial communities. Soil bacterial communities were regulated by soil properties, their changes reflected a combined effect of soil characteristics and tree species.

The orchestration of coding and non-coding genes at the CDH13 locus in coronary artery disease

Abstract Background The CDH13 locus was associated with coronary artery disease (CAD) by genome-wide association studies (GWAS). However, the causal gene(s) and underlying disease mechanisms at this locus remain undetermined. Purpose We intended to pinpoint and functionally validate the causal coding and non-coding genes at the CDH13 locus and delineate the underlying mechanisms for CAD. Methods and results We conducted the transcriptome-wide association analysis (TWAS) using nine types of CAD-relevant tissues from Stockholm-Tartu Atherosclerosis Reverse Network Engineering Task (STARNET) and Genotyped Tissue Expression (GTEx) projects and identified CDH13 (T-cadherin) and four long non-coding RNA (lncRNA) genes as candidate causal genes at The CDH13 locus. One of the antisense(AS) lncRNAs, CDH13-AS2, was predicted to bind on the 3’ untranslated region (UTR) of CDH13 using LncRRIsearch and RNAup servers. The binding interaction was validated by dCas13-medicated RNA immunoprecipitation. Weighted correlation network analysis (WGCNA) using transcriptomic data from CAD-relevant tissues of ~600 human subjects suggested a positive expression correlation between CDH13 and CDH13-AS2 in artery tissues. CRISPR/Cas9-based knockout (KO) of CDH13 or CDH13-AS2 in human umbilical vein endothelial cells (HUVECs) indicated synergistically atherogenic phenotypes, including reduced proliferation and migration, and increased apoptosis and monocyte adhesion. The data matched with the downregulation of CDH13 in artery samples of atherosclerosis patients. The cellular phenotypes were also in line with our in vivo data of Cdh13 and Apoe double-KO (Cdh13-/-/Apoe-/-) mice, which showed increased aortic lesion areas when fed on a high-fat diet compared to Apoe-/- mice. Furthermore, prediction using TargetScan, miRWalk, and scanMiRApp databases identified the potential binding of several EC-expression microRNAs (mir) on 3’UTR of CDH13. Several disease-associated variants were mapped to the predicted binding sites of a few microRNAs, including mir-let7, mir-30, and mir-125. The dual luciferase-based RNA interference (RNAi) assay confirmed the binding of mir-let7 on 3’UTR of CDH13. Transcriptional activation of CDH13-AS2 by the enzymatically inactivated Cas9 (dCas9) system diminished the binding of mir-let7 and increased CDH13 expression. Conclusions At the CDH13 locus, CDH13 and CDH13-AS2 are two causal genes, whose downregulation likely contributes to CAD. CDH13-AS2 bound and stabilized CDH13 partially by preventing mir-let7 mediated degradation of CDH13 mRNA, suggesting therapeutic potentials.