Membrane-targeted push-pull azobenzenes for the optical modulation of membrane potential

We introduce a family of membrane-targeted azobenzenes (MTs) with a push-pull character as a new tool for cell stimulation. These molecules are water soluble and spontaneously partition in the cell membrane. Upon light irradiation, they isomerize from trans to cis, changing the local charge distribution and thus stimulating the cell response. Specifically, MTs photoisomerization induces clear and reproducible depolarization. The most promising species, MTP2, was extensively studied. Time-resolved spectroscopy techniques provide insights into the excited state evolution and a complete understanding of its isomerization reaction. Molecular Dynamics simulations reveal the spontaneous and stable partitioning of the compound into the cellular membrane, without significant alterations to the bilayer thickness. MTP2 was tested in different cell types, including HEK293T cells, primary neurons, and cardiomyocytes, and a steady depolarization is always recorded. The observed membrane potential modulation in in-vitro models is attributed to the variation in membrane surface charge, resulting from the light-driven modulation of the MT dipole moment within the cell membrane. Additionally, a developed mathematical model successfully captures the temporal evolution of the membrane potential upon photostimulation. Despite being insufficient for triggering action potentials, the rapid light-induced depolarization holds potential applications, particularly in cardiac electrophysiology. Low-intensity optical stimulation with these modulators could influence cardiac electrical activity, demonstrating potential efficacy in destabilizing and terminating cardiac arrhythmias. We anticipate the MTs approach to find applications in neuroscience, biomedicine, and biophotonics, providing a tool for modulating cell physiology without genetic interventions

Leveraging Geographical Information to Strengthen People’s Engagement in Local Placemaking Processes

Participatory mapping approaches are increasingly employed in social and environmental fields of research and practice. This chapter explores how the use of geographical information can enhance people’s engagement in placemaking. Starting from theoretical concepts and experiences, the chapter explores the use of geographical information in three different contexts. The three cases all aimed to use geographical information to engage people in public participation processes, albeit with different tools and approaches and pursuing different objectives. In the province of Brindisi, a combination of a map-based questionnaire and geo-design workshops was used to bring to the fore local heritage perceptions and to involve the local community incultural landscape planning. In the case of Ancona, a web-based public participation geographic information system (PPGIS) related to abandoned buildings and public spaces was developed in a university course together with third-party associations. The objectives were to identify meaningful places and spaces and how they could be reused and regenerated and to envisage an alternative urban development. In urban Morelia, Mexico, the focus was on participatory mapping with children concerning their perceptions of their home neighbourhoods and the journeys between home and school, especially concerning risk places. The tools used were mental maps, Google Earth images and GeoODK for recording routes and places. All the case studies discuss the strengths and limitations of the various methods applied and how geographical information can involve different groups of participants in placemaking processes. Key lessons can be learnt from critically assessing these specific approaches and tools towards enhancing engagement using geographical information. Among these are: how geographical information can strengthen representation of people’s inputs and ideas; how to respond to issues of representativity and inclusion of diverse participants and their trust towards external organisers; and the necessity of clarity and self-awareness in collaboratively determining the purpose of the participatory mapping processes

Developmental validation of a multiplex qPCR assay for simultaneous quantification of nuclear and mitochondrial DNA

Quantification of human DNA is key in forensic genetics. A more accurate estimate of the amount of DNA is essential for planning and optimising genotyping assays, as is evaluating the presence of PCR inhibitory substances and DNA degradation status. Multiplex qPCR assays are helpful in forensics because they can quantify different targets simultaneously, thus saving valuable samples, time, and labour. The aim of this study was to highlight the challenges in the developmental validation of a multiplex real-time PCR assay and the drawbacks encountered in translating a previously described and validated assay (SD quants) to a different technology by modifying the dye probes and reagent mix to be used in a different instrument. We developed a TaqMan probebased multiplex qPCR using reagents and fluorescent probes adapted for the Rotor-Gene 6000 instrument (QIAGEN, Hilden, Germany). The initial assay combined two mitochondrial DNA (mtDNA) and two nuclear DNA (nDNA) targets, with amplification products of different sizes (mtDNA = 69 and 143 bp; nDNA = 71 and 181 bp), to estimate the DNA degradation status and an internal positive control (IPC) to detect potential inhibitors. During the initial testing of the assay, we observed an interaction between the 69 bp mtDNA target and the 71 bp nDNA target probe, and experiments were conducted to resolve this issue without success. We removed the small nDNA target (71 bp) and changed from a 5-plex to a 4-plex qPCR assay (qMIND). The final tetraplex assay was tested on 105 forensic samples and/or small amounts of degraded DNA, such as bones, teeth, fingernails, formalin-fixed paraffin-embedded tissues (FFPE), and hair shaft samples. The quantification results were compared with data acquired from the same samples using another commercially available quantification system commonly used in forensic laboratories. In addition, the short tandem repeat (STR) profiles were investigated to determine their correlation with the quantitative values obtained. Overall, the qPCR assay was robust and reliable for DNA quantification in samples commonly used in forensic practice

Molecularly imprinted polymers (MIPs) for SARS-CoV-2 omicron variant inhibition: An alternative approach to address the challenge of emerging zoonoses

Emerging zoonoses pose significant public health risks and necessitate rapid and effective treatment responses. This study enhances the technology for preparing Molecularly Imprinted Polymers (MIPs), which function as synthetic nanoparticles targeting SARS-CoV-2 receptor-binding domain (RBD), specifically the Omicron variant, thereby inhibiting its function. This study builds on previous findings by introducing precise adjustments in the formulation and process conditions to enhance particle stability and ensure better control over size and distribution, thereby overcoming the issues identified in earlier research. Following docking studies, imprinted nanoparticles were synthesized via inverse microemulsion polymerization and characterized in terms of size, morphology and surface charge. The selective recognition properties and ability of MIPs to obstruct the interaction between ACE2 and the RBD of SARS-CoV-2 were assessed in vitro, using Non-Imprinted Polymers (NIPs) as controls, and rebinding studies were conducted utilizing a Quartz Crystal Microbalance with Dissipation monitoring (QCM-D). The synthesized nanoparticles exhibited uniform dispersion and had a consistent diameter within the nanoscale range. MIPs demonstrated significant recognition properties and exhibited a concentrationdependent ability to reduce RBD binding to ACE2 without cytotoxic or sensitizing effects. MIPs-based platforms offer a promising alternative to natural antibodies for treating SARS-CoV-2 infections, therefore representing a versatile platform for managing emerging zoonoses

Bi-Directional Relationship Between Bile Acids (BAs) and Gut Microbiota (GM): UDCA/TUDCA, Probiotics, and Dietary Interventions in Elderly People

The gut microbiota (GM), the set of microorganisms that colonizes our intestinal tract, can undergo many changes, some of which are age related. Several studies have shown the importance of maintaining a healthy GM for a good quality of life. In the elderly, maintaining a good GM may become a real defense against infection by pathogens, such as C. difficile. In addition to the GM, bile acids (BAs) have been shown to provide an additional defense mechanism against the proliferation of pathogenic bacteria and to regulate bacterial colonization of the gut. BAs are molecules produced in the host liver and secreted with the bile into the digestive tract, and they are necessary for the digestion of dietary lipids. In the gut, host-produced BAs are metabolized by commensal bacteria to secondary BAs. In general GM and host organisms interact in many ways. This review examines the relationship between GM, BAs, aging, and possible new approaches such as dietary interventions, administration of ursodesoxycholic acid/tauroursodesoxycholic acid (UDCA/TUDCA), and probiotics to enrich the microbial consortia of the GM in the elderly and achieve a eubiotic state necessary for maintaining good health. The presence of Firmicutes and Actinobacteria together with adequate levels of secondary BAs would provide protection and improve the frailty state in the elderly. In fact, an increase in secondary BAs has been observed in centenarians who have reached old age without serious health issues, which may justify their active role in achieving longevity

Genome-Wide Association Integrating a Transcriptomic Meta-Analysis Suggests That Genes Related to Fat Deposition and Muscle Development Are Closely Associated with Growth in Huaxi Cattle

Growth traits are among the most important economic phenotypes targeted in the genetic improvement of beef cattle. To understand the genetic basis of growth traits in Huaxi cattle, we performed a genome-wide association study (GWAS) on body weight, eye muscle area, and back fat thickness across five developmental stages in a population of 202 Huaxi cattle. Additionally, publicly available RNA-seq data from the longissimus dorsi muscle of both young and adult cattle were analyzed to identify key genes and genetic markers associated with growth in Huaxi cattle. In total, 7.19 million high-quality variant loci (SNPs and INDELs) were identified across all samples. In the GWAS, the three multilocus models (FarmCPU, MLMM, and BLINK) outperformed the conventional single-locus models (CMLM, GLM, and MLM). Consequently, GWAS analysis was conducted using multilocus models, which identified 99 variant loci significantly associated with growth traits and annotated a total of 83 candidate genes (CDGs). Additionally, 23 of the 83 CDGs overlapped with significantly differentially expressed genes identified from public RNA-seq datasets of longissimus dorsi muscle between young and adult cattle. Furthermore, gene functional enrichment (KEGG and GO) analyses revealed that over 30% of the pathways and GO terms were associated with muscle development and fat deposition, crucial factors for beef production. Specifically, key genes identified included MGLL, SGMS1, SNX29 and AKAP6, which are implicated in lipid metabolism, adipogenesis, and muscle growth. In summary, this study provides new insights into the genetic mechanisms underlying growth traits in Huaxi cattle and presents promising markers for future breeding improvements

Effects of Mandibular Advancement Devices in Patients Affected by OSA and Bruxism: A Finite Element Study

Obstructive Sleep Apnea (OSA) is a common respiratory disorder char-acterised by episodes of complete or partial obstruction of the upper airways dur-ing sleep. Nearly half of adults with OSA have comorbid Sleep Bruxism (SB), a condition in which a person grinds, gnashes, or clenches the teeth during sleep. However, sparse literature exists about the Finite Element Analysis (FEA) of mandibular advancement devices (MADs) for OSA therapy under a biting load. This manuscript presents a FEA to evaluate the effects of MADs (i.e., stress and displacement on periodontal ligaments and teeth) under a typical clenching load due to SB (i.e., 650 N), compared to the only OSA condition (baseline). Findings have demonstrated that the teeth stress and displacement are strictly related to the load magnitude and the MAD geometry (plateaux and relative position between the splints). Indeed, the applied load shifts the maximum values of stress and displacement from the MAD connecting sites to the contact area of the splints, increasing the stress in the anterior region of the maxilla. Despite this study being developed on one single patient and one single MAD model, this preliminary investigation can provide useful insights into the consequences of MAD usage in people affected by SB

Nonlinear vibrations of kinematically exact curved beams

The nonlinear oscillations of a kinematically exact curved beam are investigated by means of the multiple time scale method applied directly to partial differential equations of motion. A linear constitutive behaviour is assumed, and the bending strain is the (change of) mechanical curvature. A dependence of natural frequencies ωi and nonlinear correction coefficients ncci (describing the nonlinear behaviour of the beam) on the initial curvature α is investigated for the first six modes for a case of hinged–hinged boundary conditions. The occurrence of internal resonances is discussed, and a complex behaviour of the functions ncci(α) is illustrated in detail. A comparison is made with the results obtained by the single mode Galerkin approximation, showing that the latter yields incorrect results. Finally, the analytical solution is validated by comparing it with numerical simulations obtained by the finite element method

Endothelial cell alterations in capillaries of adipose tissue from patients affected by lipedema

Objective: This study aimed to evaluate adipose tissue of lipedema patients. Methods: Gluteo-femoral (affected area) and interscapular (nonaffected area) adipose tissue from 10 lean patients affected by lipedema stage 1 to 2 was studied and compared with tissue from 10 patients with obesity and 12 lean patients. Results: The main features were alterations of capillaries with wall thickening (p ≤ 0.0001), endothelial and pericyte hyperplasia (p = 0.03 and p = 0.004), hypodense areas in basal membrane, and endothelial degeneration with exfoliation of degenerated cells into the capillary lumen. Adipocytes were larger (hypertrophic) in affected (P ≤ 0.0001) and nonaffected (p = 0.0003) areas compared with those with obesity and who were lean (both p ≤ 0.0001). Frequently the cytoplasm of adipocytes contained massive deposition of calcium crystals as revealed by Von Kossa staining (p = 0.023) and electron microscopy. CD68 immunoreactive macrophages were more abundant in affected areas (p = 0.005), and their number was similar to that found in fat from patients with obesity (p = 0.17). Despite adipocyte hypertrophy and inflammation, lack of the healthy marker perilipin-1 and the presence of crown-like structures were only rarely seen, while they were quite frequent in patients with obesity. Conclusions: Our data support the idea that cell alterations happen in the early stages of adipocyte development (endothelium/pericyte) in the adipose organ of women affected by lipedema

Towards Sustainable Furniture: Evaluating Environmental Impact Perception and Tools for Circular Design

The increasing awareness of the necessity for a circular approach in both the government and private sectors has led to the adoption of circular busi-ness models aimed at enhancing sustainability efforts and reducing waste pro-duction. Despite this recognition, there remains a lack of empirical research on how companies evaluate their environmental impacts to forecast the consequences of applying circular models. This study aims to address this gap by examining the environmental impact perception and diffusion of methods and tools in the furniture sector. Through literature review and questionnaire surveys, the study identifies key concepts and highlights varying levels of commitment to sustain-ability across different company sizes. The obtained evidence underscores the importance of simplified Life Cycle Assessment (LCA) tools in guiding designers towards sustainable product choices, especially in small companies with limited environmental sustainability culture. The outcome emphasizes the need for a com-prehensive understanding of environmental impacts for the effective adoption of circular business models and provides insights for developing tools to support eco-design and circular strategies in the furniture sector