Matrix Interactions Research Articles

Anoikis in phenotypic reprogramming of the prostate tumor microenvironment.

Prostate cancer is one of the most common malignancies in males wherein 1 in 8 men are diagnosed with this disease in their lifetime. The urgency to find novel therapeutic interventions is associated with high treatment resistance and mortality rates associated with castration-resistant prostate cancer. Anoikis is an apoptotic phenomenon for normal epithelial or endothelial cells that have lost their attachment to the extracellular matrix (ECM). Tumor cells that lose their connection to the ECM can die via apoptosis or survive via anoikis resistance and thus escaping to distant organs for metastatic progression. This review discusses the recent advances made in our understanding of the signaling effectors of anoikis in prostate cancer and the approaches to translate these mechanistic insights into therapeutic benefits for reducing lethal disease outcomes (by overcoming anoikis resistance). The prostate tumor microenvironment is a highly dynamic landscape wherein the balance between androgen signaling, cell lineage changes, epithelial-mesenchymal transition (EMT), extracellular matrix interactions, actin cytoskeleton remodeling as well as metabolic changes, confer anoikis resistance and metastatic spread. Thus, these mechanisms also offer unique molecular treatment signatures, exploitation of which can prime prostate tumors to anoikis induction with a high translational significance.

Abstract 3348: Unraveling new strategies for early detection of invasive cancer cells: A special focus on the nuclear and cytoskeletal architecture

Abstract Introduction: Cancer invasion is a hallmark of cancer progression and the leading cause of cancer mortality worldwide. However, the identification of invasive cancer cells at an early stage remains challenging. Therefore, there is an urgent need to develop novel biomarkers and innovative strategies to successfully tackle cancer invasion. It is well established that the structure and organization of the nucleus and the cytoskeleton are dynamically orchestrated during many cellular processes, including cancer invasion. Thus, in this study, we investigated fine-tuned nuclear and cytoskeletal patterns associated with invasion, taking into consideration the cell-extracellular matrix (ECM) interaction. Methods: In this study, we took advantage of an ECM Array platform (MicroMatrix 36, MicroStem) and a panel of invasive and non-invasive gastric cancer cells. For validation purposes, we assessed cells transfected with wild type or mutant E-cadherin, which lead to distinct adhesion and invasion phenotypes. A comprehensive analysis of nuclei and cytoskeletal architectural features was performed. Nuclei area, intensity, perimeter, eccentricity, circularity, solidity and entropy were evaluated. Notably, a new computational pipeline was developed to characterize cytoskeletal structures, namely orientation, compactness, radiality, bundling, parallelness, morphology, among others. Results and Discussion: Our results have shown that the invasive abilities of cancer cells are associated with unique nuclear and cytoskeletal modifications. Moreover, we verified that invasive cells adhere preferentially to specific ECM components. Interestingly, we could observe an association between nuclear phenotypes and increased attachment abilities, suggesting that the ECM may modulate nuclear morphometrics thus affecting the invasive behavior of cancer cells. Of note, cell lines harboring E-cadherin mutations corroborated the existence of a nuclear signature associated with invasive potential. Remarkably, the proposed computational framework has shown that the microtubules of cells with disrupted E-cadherin are shorter, have uniform length patterns and are more compactly distributed as compared with cells with wild-type E-cadherin. Conclusion: Overall, this study demonstrates that the use of nuclear and cytoskeletal architectural features could provide an efficient strategy to identify cells with invasive potential. Further investigation and integration of this approach with other cellular properties could have an impact in diagnosis, prognosis and therapeutic strategies. Citation Format: Maria Sofia Fernandes, Joana Figueiredo, Diogo Vieira, Soraia Melo, Ana Margarida Moreira, João Miguel Sanches, Raquel Seruca. Unraveling new strategies for early detection of invasive cancer cells: A special focus on the nuclear and cytoskeletal architecture [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3348.

Abstract 4557: 3D-EXpress platform utilizing tumoroids from patients with MSS and MSI-H tumors allows rapid assessment of anti-tumor activity of immune checkpoint inhibitors and development of clinically relevant biomarkers of treatment response

Abstract Introduction: Recent studies showed that patients with deficient mismatch repair(dMMR)/microsatellite instability-high (MSI-H) tumors, have higher sensitivity to immune checkpointinhibitors (ICIs) compared to patients with microsatellite-stable (MSS)/microsatellite instability-low(MSI-L) tumors. However, the mechanisms of treatment responsiveness and resistance is not wellunderstood. Here, we used a novel 3D-EXpress ex vivo fresh patient tumoroid platform to assess theefficacy of nivolumab/ipilimumab combination therapy in tumors with known MSS/MSI-H status andperformed correlative studies. Materials and Methods: All tumor samples were obtained with patient consent and relevant IRBapproval. Tumoroids measuring 150 µm in size retaining tumor cell heterogeneity, tumor-residentimmune cells, stromal components, and cell-extracellular matrix interaction were prepared frompatients with endometrial and colorectal tumors among others. Tumoroid aliquots were cryopreservedin Nilogen’s tumoroid Biorepository for future studies. For the 3D-EXpress studies cryopreservedtumoroids were selected based on MSS/MSI-H status and treated ex vivo with nivolumab andipilimumab alone and in combinations for 72h. Tumor responses to treatments were evaluated by aproprietary tumor cell killing assay and changes in tumor immune microenvironment. Furthermore,tumor PD-L1 expression levels were analyzed on the associated TMA slides. Results: Treatment-induced tumor cell killing activity in intact tumoroids was assessed by a 3D high-content confocal imaging technique using a proprietary algorithm for data analysis. The impact of exvivo treatment by nivolumab and/or ipilimumab on tumor resident immune cell populations wasmonitored by a multiplex cytokine release assay analyzing release of GM-CSF, sCD137, IFNγ, sFas, sFasL,Granzyme A, Granzyme B, IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, MIP-1α, MIP-1β, TNF-α, and Perforin usingculture supernatants isolated from treated tumoroid samples. Based on the ex vivo responses tumorswere assigned to treatment sensitive and resistant groups and correlative analyses were performed withindividual tumors’ innate and adaptive immune cell populations detected by a 21-color flow cytometrypanel, in addition to tumor MSS/MSI-H and p53 status, and detailed clinicopathologic data readilyavailable for the tumors cryopreserved in the Biorepository. Conclusion: The 3D-Express platform, using cryopreserved 3D tumoroids with intact TME is an effectivetool for the assessment of rational combinations which may prove relevant in the treatment of solidtumors. Furthermore, we believe this platform is a useful tool for the pre-clinical assessment of specifictherapeutic regimens designed for individualized patient care. Citation Format: Brittney Ruedlinger, Seth Currlin, Sharon Camacho, Alliyah Humphrey, Jared Ehrhart, Soner Altiok. 3D-EXpress platform utilizing tumoroids from patients with MSS and MSI-H tumors allows rapid assessment of anti-tumor activity of immune checkpoint inhibitors and development of clinically relevant biomarkers of treatment response. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4557.

Abstract 2209: A proteomic-based prognostic signature of pancreatic adenocarcinoma

Abstract Introduction: Pancreatic cancer has a very poor prognosis, with no established prognostic biomarkers in clinical use. This project aims to identify a prognostic proteomic-based signature for pancreatic adenocarcinomas. Methods: Fresh frozen tumors and matched normal samples from 125 patients were prepared for proteomic analyses using data-independent acquisition mass spectrometry (DIA-MS). Differential expression analyses were conducted on the normalized protein matrix to identify the top differentially expressed proteins (DEP) within the tumor samples. DEP were subjected to crosstalk and pathway enrichment analysis (PEA). Survival analysis based on initial univariate and subsequent 100 runs of multivariate Cox regression with Least Absolute Shrinkage and Selection Operator (LASSO) was performed to obtain a reduced list of candidate proteins associated with Overall Survival (OS). The proteins that appeared in greater than 95% of the LASSO runs were then used in a multivariate Cox model with recursive feature selection, which yielded the final 29 proteins. A risk score was built from the final 29 proteins. Consensus clustering was performed on the median absolute deviation-based top 20% highly variable proteins in tumor samples to identify proteomic-based subtypes. Results: Proteomic analyses revealed 5614 proteins identified from 599 sample runs. Differential expression analyses revealed 398 DEP in tumor samples (FDR-adjusted p-value <0.05, and |logFC|>1). PEA showed that these proteins were related to focal adhesion, extracellular matrix interaction (ECM), angiogenesis, and PI3K signaling pathways. A total of 803 proteins were significantly associated with OS in a univariate Cox regression analysis (p<0.05). PEA on the top 200 proteins associated with poorer OS revealed pathways related to focal adhesion, PI3K signaling, ECM and hypoxia-induced factor-1. Using LASSO multivariate Cox regression modeling, a 29-protein signature was identified, from which a risk score was calculated that dichotomized patients into high- and low-risk groups in terms of OS (Hazard ratio (HR) 2.8, 95% Confidence Interval (CI) [2.3, 3.3], concordance index of 0.91). This risk score was also prognostic for recurrence and three-year survival (both p<0.0001). A multivariate Cox regression model adjusted for other clinical variables revealed a significant association of the risk score with OS (HR 2.91, 95% CI [2.4, 3.5], p<0.001) while maintaining the concordance index (0.907). Consensus clustering analyses revealed four proteomic-based clusters, with cluster 3 showing the worst OS (p<0.001), independent of other clinical variables. PEA on the DEP within cluster 3 showed upregulation of proteins related to cell adhesion, angiogenesis, and immune-related pathways. Conclusion: A 29-protein signature identified a sub-group of patients with pancreatic adenocarcinoma with a poorer prognosis independent of clinical variables. Citation Format: Adel T. Aref, AKM Azad, Asim Anees, Mohashin Pathan, Jason Grealey, Daniela-Lee Smith, Erin M. Humphries, Daniel Bucio-Noble, Jennifer M. Koh, Erin Sykes, Steven G. Williams, Ruth Lyons, Natasha Lucas, Dylan Xavier, Sumit Sahni, Anubhav Mittal, Jaswinder S. Samra, John V. Pearson, Nicola Waddell, Peter G. Hains, Phil J. Robinson, Qing Zhong, Roger R. Reddel, Anthony J. Gill. A proteomic-based prognostic signature of pancreatic adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2209.

A2AR as a Prognostic Marker and a Potential Immunotherapy Target in Human Glioma.

Gliomas are considered one of the most malignant tumors in the body. The immune system has the ability to control the initiation and development of tumors, including gliomas. Thus, immune cells find themselves controlled by various molecular pathways, inhibiting their activation, such as the immunosuppressive adenosine 2A receptor (A2AR). Our objective was to establish the expression profile and role of A2AR at the transcriptomic level, using real-time RT-PCR in Moroccan glioma patients, in addition to TCGA and CGGA cohorts. The real-time RT-PCR results in Moroccan patients showed that high expression of this gene was associated with poor survival in males. Our study on the CGGA cohort corroborated these results. In addition, there was a positive association of A2AR with T-cell exhaustion genes. A2AR also correlated strongly with genes that are primarily enriched in focal adhesion and extracellular matrix interactions, inducing epithelial mesenchymal transition, angiogenesis, and glioma growth. However, in the TCGA cohort, the A2AR showed results that were different from the two previously examined cohorts. In fact, this gene was instead linked to a good prognosis in patients with the astrocytoma histological type. The correlation and enrichment results reinforced the prognostic role of A2AR in this TCGA cohort, in which its high expression was shown to be related to lymphocyte differentiation and a successful cytolytic response, suggesting a more efficient anti-tumor immune response. Correlations and differential analyses based on A2AR gene expression, to understand the cause of the association of this gene with two different prognoses (CGGA males and TCGA Astrocytoma), showed that the overexpression of A2AR in Chinese male patients could be associated with the overexpression of extracellular adenosine, which binds to A2AR to induce immunosuppression and consequently a poor prognosis. However, in the second group (TCGA astrocytomas), the overexpression of the gene could be associated with an adenosine deficiency, and therefore this receptor does not undergo activation. The absence of A2AR activation in these patients may have protected them from immunosuppression, which could reflect the good prognosis. A2AR can be considered a promising therapeutic target in male CGGA and Moroccan patients with gliomas.

The Mechanism of the Anti-Cardiac Hypertrophy Effect of Glycyrrhizic Acid Is Related to Reducing STIM1-Dependent Store-Operated Calcium Entry

We explored the anti-cardiac hypertrophy mechanism of glycyrrhizic acid from the perspective of calcium regulation under pathological conditions. For this purpose, we used a rat model of myocardial hypertrophy induced by pressure overload. The effect of glycyrrhizic acid on BP was measured non-invasively with a sphygmomanometer and recorded in PC. In rats with modeled cardiac hypertrophy, the effect of GA on expression of type 1 matrix interaction molecules was determined in horizontal tissues and cultured cardiomyocytes of the left ventricle. The laser confocal microscopy and calcium ion probe Fluo-4 AM were used to assess the effect of glycyrrhizic acid on stromal interaction molecule 1 (STIM1)-dependent store-operated calcium entry in cultured cardiomyocytes derived from the hypertrophic myocardium. Glycyrrhizic acid exerted the anti-hypertrophic effect in rats with hypertrophic myocardium by down-regulating STIM1 protein expression and reducing the intensity of STIM1-dependent store-operated calcium entry.

Application of microfluidic chips in the simulation of the urinary system microenvironment.

The urinary system, comprising the kidneys, ureters, bladder, and urethra, has a unique mechanical and fluid microenvironment, which is essential to the urinary system growth and development. Microfluidic models, based on micromachining and tissue engineering technology, can integrate pathophysiological characteristics, maintain cell-cell and cell-extracellular matrix interactions, and accurately simulate the vital characteristics of human tissue microenvironments. Additionally, these models facilitate improved visualization and integration and meet the requirements of the laminar flow environment of the urinary system. However, several challenges continue to impede the development of a tissue microenvironment with controllable conditions closely resemble physiological conditions. In this review, we describe the biochemical and physical microenvironment of the urinary system and explore the feasibility of microfluidic technology in simulating the urinary microenvironment and pathophysiological characteristics in vitro. Moreover, we summarize the current research progress on adapting microfluidic chips for constructing the urinary microenvironment. Finally, we discuss the current challenges and suggest directions for future development and application of microfluidic technology in constructing the urinary microenvironment in vitro.

Cell-matrix interactions control biliary organoid polarity, architecture, and differentiation.

Cholangiopathies are an important cause of morbidity and mortality. Their pathogenesis and treatment remain unclear in part because of the lack of disease models relevant to humans. Three-dimensional biliary organoids hold great promise; however, the inaccessibility of their apical pole and the presence of extracellular matrix (ECM) limits their application. We hypothesized that signals coming from the extracellular matrix regulate organoids' 3-dimensional architecture and could be manipulated to generate novel organotypic culture systems. Biliary organoids were generated from human livers and grown embedded into Culturex Basement Membrane Extract as spheroids around an internal lumen (EMB). When removed from the EMC, biliary organoids revert their polarity and expose the apical membrane on the outside (AOOs). Functional, immunohistochemical, and transmission electron microscope studies, along with bulk and single-cell transcriptomic, demonstrate that AOOs are less heterogeneous and show increased biliary differentiation and decreased expression of stem cell features. AOOs transport bile acids and have competent tight junctions. When cocultured with liver pathogenic bacteria (Enterococcus spp.), AOOs secrete a range of proinflammatory chemokines (ie, MCP1, IL8, CCL20, and IP-10). Transcriptomic analysis and treatment with a beta-1-integrin blocking antibody identified beta-1-integrin signaling as a sensor of the cell-extracellular matrix interaction and a determinant of organoid polarity. This novel organoid model can be used to study bile transport, interactions with pathobionts, epithelial permeability, cross talk with other liver and immune cell types, and the effect of matrix changes on the biliary epithelium and obtain key insights into the pathobiology of cholangiopathies.

Mineralization of Bone Extracellular Matrix-like Scaffolds Fabricated as Silk Sericin-Functionalized Dense Collagen-Fibrin Hybrid Hydrogels.

The design of hydrogels that combine both the biochemical cues needed to direct seeded cellular functions and mineralization to provide the structural and mechanical properties approaching those of mineralized native bone extracellular matrix (ECM) represents a significant challenge in bone tissue engineering. While fibrous hydrogels constituting of collagen or fibrin (and their hybrids) can be considered as scaffolds that mimic to some degree native bone ECM, their insufficient mechanical properties limit their application. In the present study, an automated gel aspiration-ejection (automated GAE) method was used to generate collagen-fibrin hybrid gel scaffolds with micro-architectures and mechanical properties approaching those of native bone ECM. Moreover, the functionalization of these hybrid scaffolds with negatively charged silk sericin accelerated their mineralization under acellular conditions in simulated body fluid and modulated the proliferation and osteoblastic differentiation of seeded MC3T3-E1 pre-osteoblastic cells. In the latter case, alkaline phosphatase activity measurements indicated that the hybrid gel scaffolds with seeded cells showed accelerated osteoblastic differentiation, which in turn led to increased matrix mineralization. In summary, the design of dense collagen-fibrin hybrid gels through an automated GAE process can provide a route to tailoring specific biochemical and mechanical properties to different types of bone ECM-like scaffolds, and can provide a model to better understand cell-matrix interactions in vitro for bioengineering purposes.

Reprogramming of the Genome-Wide DNA Methylation Landscape in Three-Dimensional Cancer Cell Cultures

During the last century, 2D cell cultures have been the tool most widely used to study cancer biology, drug discovery, genomics, and the regulation of gene expression at genetic/epigenetic levels. However, this experimental approach has limitations in faithfully recreating the microenvironment and cellular processes occurring in tumors. For these reasons, 3D cell cultures have recently been implemented to optimize the conditions that better recreate the biological and molecular features of tumors, including cell-cell and cell-extracellular matrix (ECM) interactions, growth kinetics, metabolic activities, and the development of gradients in the cellular microenvironment affecting the availability of oxygen and nutrients. In this sense, tumor cells receive stimuli from the local environment, resulting in significant changes in their signaling pathways, gene expression, and transcriptional and epigenetic patterns. In this review, we discuss how different types of 3D cell culture models can be applied to characterize the epigenetic footprints of cancer cell lines, emphasizing that DNA methylation patterns play an essential role in the emergence and development of cancer. However, how 3D cancer cell cultures remodel the epigenetic programs is poorly understood, with very few studies in this emerging topic. Here, we have summarized the studies on the reprogramming of the epigenetic landscape of DNA methylation during tumorigenesis and discuss how it may be affected by microenvironmental factors, specifically in 3D cell cultures.

A unified approach to designing sequence-based personalized food recommendation systems: tackling dynamic user behaviors

The recommender system (RS) is a well-known practical application of the state-of-the-art information filtering and machine learning technologies. Traditional recommendation approaches, including collaborative and content-based filtering techniques, have been widely employed to provide suggestions in RSs, where the user-item interaction matrix is the primary data source. In many application domains, interactions between users and items are more likely to be dynamic rather than static, and thus dynamic user behaviors should be taken into account when solving recommendation tasks in order to provide more accurate suggestions. In this work, we consider the sequentially ordered information from user-item interactions in the RSs where a sequence-based recommendation model is put forward with applications to the food recommendation scenario. Furthermore, the long short-term memory (LSTM) network is employed as the building block to establish such a recommendation model, and a collaborative filtering unit is adopted to make personalized food recommendation. The proposed LSTM-based RS is successfully applied to a real-world food recommendation data set. Experimental results demonstrate that the developed method outperforms some currently popular RSs in terms of precision, recall, mean average precision and mean reciprocal rank in food recommendation.

Cellulose nanocrystal functionalized aramid nanofiber reinforced epoxy nanocomposites with high strength and toughness

The mechanical properties of polymer nanocomposites can be improved by incorporating various types of nanofillers. The hybridization of nanofillers through covalent linkages between nanofillers with different dimensions and morphology can further increase the properties of nanocomposites. In this work, aramid nanofibers (ANFs) are modified using chlorinated cellulose nanocrystals (CNCs) and functionalized with 3-glycidoxypropyltrimethoxysilane to improve the chemical and mechanical interaction in an epoxy matrix. The integration of CNC functionalized ANFs (fACs) in the epoxy matrix simultaneously improves Young’s modulus, tensile strength, fracture properties, and viscoelastic properties. The test results show that 1.5 wt% fAC reinforced epoxy nanocomposites improve Young’s modulus and tensile strength by 15.1% and 10.1%, respectively, and also exhibit 2.5 times higher fracture toughness compared to the reference epoxy resin. Moreover, the glass transition temperature and storage modulus are found to increase when fACs are incorporated. Thus, this study demonstrates that the enhanced chemical and mechanical interaction by the CNC functionalization on the ANFs can further improve the static and dynamic mechanical properties of polymer nanocomposites.

Pericytes: The lung-forgotten cell type.

Pericytes are a heterogeneous population of mesenchymal cells located on the abluminal surface of microvessels, where they provide structural and biochemical support. Pericytes have been implicated in numerous lung diseases including pulmonary arterial hypertension (PAH) and allergic asthma due to their ability to differentiate into scar-forming myofibroblasts, leading to collagen deposition and matrix remodelling and thus driving tissue fibrosis. Pericyte-extracellular matrix interactions as well as other biochemical cues play crucial roles in these processes. In this review, we give an overview of lung pericytes, the key pro-fibrotic mediators they interact with, and detail recent advances in preclinical studies on how pericytes are disrupted and contribute to lung diseases including PAH, allergic asthma, and chronic obstructive pulmonary disease (COPD). Several recent studies using mouse models of PAH have demonstrated that pericytes contribute to these pathological events; efforts are currently underway to mitigate pericyte dysfunction in PAH by targeting the TGF-β, CXCR7, and CXCR4 signalling pathways. In allergic asthma, the dissociation of pericytes from the endothelium of blood vessels and their migration towards inflamed areas of the airway contribute to the characteristic airway remodelling observed in allergic asthma. Although several factors have been suggested to influence this migration such as TGF-β, IL-4, IL-13, and periostin, recent evidence points to the CXCL12/CXCR4 pathway as a potential therapeutic target. Pericytes might also play an essential role in lung dysfunction in response to ageing, as they are responsive to environmental risk factors such as cigarette smoke and air pollutants, which are the main drivers of COPD. However, there is currently no direct evidence delineating the contribution of pericytes to COPD pathology. Although there is a lack of human clinical data, the recent available evidence derived from in vitro and animal-based models shows that pericytes play important roles in the initiation and maintenance of chronic lung diseases and are amenable to pharmacological interventions. Therefore, further studies in this field are required to elucidate if targeting pericytes can treat lung diseases.

Identifying the Landscape Security Pattern in Karst Rocky Desertification Area Based on Ecosystem Services and Ecological Sensitivity: A Case Study of Guanling County, Guizhou Province

Ecological environmental security in karst areas is an issue of global concern. Identifying the ecological landscape security pattern (ELSP) is key to promoting environmental protection and alleviating the land development and utilization impacts. Ecological sources (ESs) and ecological corridors (ECs) are important bases for constructing an ELSP. We used five influencing factors (land use type, digital elevation model (DEM), rocky desertification degree, normalized difference vegetation index (NDVI) and slope) to obtain the distribution of the importance and sensitivity values of ecosystem services in Guanling County, Guizhou Province. The probability of the connectivity index (PC) was calculated, and the ES was extracted by combining the importance of ecosystem services, ecological sensitivity, and landscape connectivity. According to the topographic and geomorphological characteristics of Guanling County, seven indicators of elevation, slope, landscape type, degree of stone desertification, distance from rivers, distance from settlements, and distance from roads were selected as resistance factors for the outward expansion of the ESs to calculate the comprehensive resistance surface of Guanling County. Based on the gravity model, an interaction matrix between 10 ESs was constructed, and the magnitude of the interaction forces between the source sites was quantitatively evaluated to distinguish the important ECs and general ECs. The study showed that the total length of the ECs in Guanling County was 509.78 km, and the core area of Guanling County was large, accounting for 65.73% of the ecological landscape area. By assessing the importance of ecosystem services, ecological sensitivity, and landscape connectivity, 10 ES and 45 EC were obtained based on ArcGIS10.8, which constituted the landscape security pattern of Guanling County by ESs and ECs. Suggestions were proposed for a planning layout that will benefit the ecological restoration of Guanling County and environmental protection of the karst region according to the study area characteristics.

Application of Amyloid-Based Hybrid Membranes in Drug Delivery

The properties of amyloid fibrils, e.g., unique structural characteristics and superior biocompatibility, make them a promising vehicle for drug delivery. Here, carboxymethyl cellulose (CMC) and whey protein isolate amyloid fibril (WPI-AF) were used to synthesize amyloid-based hybrid membranes as vehicles for the delivery of cationic and hydrophobic drugs (e.g., methylene blue (MB) and riboflavin (RF)). The CMC/WPI-AF membranes were synthesized via chemical crosslinking coupled with phase inversion. The zeta potential and scanning electron microscopy results revealed a negative charge and a pleated surface microstructure with a high content of WPI-AF. FTIR analysis showed that the CMC and WPI-AF were cross-linked via glutaraldehyde and the interacting forces between membrane and MB or RF was found to be electrostatic interaction and hydrogen bonding, respectively. Next, the in vitro drug release from membranes was monitored using UV-vis spectrophotometry. Additionally, two empirical models were used to analyze the drug release data and relevant rate constant and parameters were determined accordingly. Moreover, our results indicated that in vitro drug release rates depended on the drug-matrix interactions and transport mechanism, which could be controlled by altering the WPI-AF content in membrane. This research provides an excellent example of utilizing two-dimensional amyloid-based materials for drug delivery.

Current possibilities and future opportunities provided by three-dimensional lung ECM-derived hydrogels.

Disruption of the complex interplay between cells and extracellular matrix (ECM), the scaffold that provides support, biochemical and biomechanical cues, is emerging as a key element underlying lung diseases. We readily acknowledge that the lung is a flexible, relatively soft tissue that is three dimensional (3D) in structure, hence a need exists to develop in vitro model systems that reflect these properties. Lung ECM-derived hydrogels have recently emerged as a model system that mimics native lung physiology; they contain most of the plethora of biochemical components in native lung, as well as reflecting the biomechanics of native tissue. Research investigating the contribution of cell:matrix interactions to acute and chronic lung diseases has begun adopting these models but has yet to harness their full potential. This perspective article provides insight about the latest advances in the development, modification, characterization and utilization of lung ECM-derived hydrogels. We highlight some opportunities for expanding research incorporating lung ECM-derived hydrogels and potential improvements for the current approaches. Expanding the capabilities of investigations using lung ECM-derived hydrogels is positioned at a cross roads of disciplines, the path to new and innovative strategies for unravelling disease underlying mechanisms will benefit greatly from interdisciplinary approaches. While challenges need to be addressed before the maximum potential can be unlocked, with the rapid pace at which this field is evolving, we are close to a future where faster, more efficient and safer drug development targeting the disrupted 3D microenvironment is possible using lung ECM-derived hydrogels.

Multi-level climate governance: examining impacts and interactions between national and sub-national emissions mitigation policy mixes in Canada

ABSTRACT Jurisdictions use an assortment of policies to reduce greenhouse gas emissions. Climate policy mixes have often evolved through the ad hoc layering of new policies onto an existing policy mix, rather than deliberate design of a complete policy portfolio. This can lead to unanticipated interactions between policies which can support or undermine policy objectives and is further complicated where climate policy is implemented at multiple jurisdictional levels. In the context of Canada and its four most populous provinces, we examine the development of climate policy mixes across jurisdictional levels between 2000 and 2020 and evaluate policy interactions. We develop an inventory of 184 climate policies, and examine each in terms of instrument type, implementation timing, technological specificity, and expected abatement. We evaluate interactions between overlapping policies both within jurisdictional levels (horizontal) and across jurisdictional levels (vertical) for their impact on emissions abatement using a policy coherence analysis framework. We find that subsidies and R&D funding were the most abundant policies (58%), although pricing and flexible regulation are expected to achieve the most abatement. Sub-national jurisdictions have often acted as policy pilots preceding federal policy implementation. We evaluate 356 policy interactions and find 74% are consistent in adding abatement. Less than 8% have a negative impact by reducing abatement, although vertical interactions between federal and provincial policies were more often negative (11%) than horizontal interactions at the federal (<3%) or provincial (<2%) levels. Although the impact of many interactions is unknown (13%), we generate interaction matrices as a foundational roadmap for future research, and for policy-makers to consider potential interactions when designing and assessing policy effectiveness. Key policy insights Climate policy mixes have expanded and diversified over the period 2000–2020 across jurisdictions in Canada. Sub-national jurisdictions have often acted as policy ‘pilots’ by implementing policy before the adoption of similar national level policy. Climate mitigation policy interactions are predominantly supportive toward achieving additional emissions abatement. Vertical interactions between federal and provincial policy can undermine the additionality of policy effort by sub-national jurisdictions. These findings emphasize the need for better coordination in climate policy mix design between national and sub-national jurisdictions.

Emotional Value in Online Education: A Framework for Service Touchpoint Assessment

To enhance the online education service experience, the emotional valence of the user was studied as an evaluation variable, and both qualitative and quantitative research were used to find how to evaluate online education service touchpoints. First, deconstruct the system service interface with the interactive touchpoint matrix, set service evaluation indicators from four aspects, visual guidance, learning resources, after-class evaluation, and interactive feedback, and build an online education service touchpoint evaluation system. Secondly, using Tencent Classroom as the target of research, an online education service rating experiment is created based on the two dimensions of emotional valence and perceptual cognition. With the aid of a questionnaire survey and analytic hierarchy process (AHP), a multidimensional evaluation of online education service touchpoints is accomplished using the learners’ emotional enjoyment, activation, dominance, touchpoint satisfaction, and importance as measuring indicators. Finally, concluding the assessment and optimization of online education service touchpoints, the evaluation data for the service are combined, and the evaluation results are generated using visual design. This study includes successful strategies and practical recommendations for boosting interest in e-learning services, user initiative, and excitement for learning.

Flavour them up! Exploring the challenges of flavoured plant‐based foods

AbstractFood flavourings are often added to enhance overall flavour experience during food consumption and their use in plant‐based food analogues is crucial. The flavour sensory perception is strongly mediated by flavour–food matrix interactions. Aroma molecules establish chemical and physical bonds with lipids and carbohydrates, but proteins play a pivotal role having a strong flavour binding minimizing aroma release and quenching flavour perception. Consequently, final food quality is reduced and so is consumer acceptance. Depending on the chemical structure of the flavour and the type of protein involved, the strength of the flavour–protein binding can vary. It is of great importance to understand if this interaction remains under dynamic conditions such as the oral processing during food consumption. This review aims to gain insights into the influence of the chemical structure and physicochemical features of both proteins and flavours on the binding mechanism. Moreover, the potentiality of coupling in vivo instrumental analysis with sensory methods to study flavour release in real time is explored. Elucidating the drivers of flavour interaction with, and release from, the food matrix is essential to develop flavour solutions for the new generation of plant‐based food products.

Evaluating fracture parameters of basalt fiber reinforced and pozzolana slurry modified recycled concrete produced from waste

Recycling of construction and demolition waste provides one of the most effective paths for sustainable development, valorising recycled concrete, and environment protection. Unfortunately, due to substandard concrete matrix quality, lower strength and fracture parameters the practical applications of recycled concrete still remain the foremost concerns. To overcome these issues, a new effective strength enhancing approach was investigated in this study. More explicitly, two types of recycled concrete consisting of 20% and 100% of pozzolana slurry encrusted recycled aggregate and strengthened with various dosages of basalt fiber (BF) were evaluated in terms of fracture properties, namely compressive strength, fracture energy, fracture toughness, flexural strength, and elastic modulus. In addition, a digital image correlation (DIC) analysis was established to evaluate the strain contour variation while the microstructure study was conducted to inspect the concrete matrix quality. Experimental result revealed improvement in the ultimate load capacity as well as the overall load-CMOD response of BF modified formulations. For 20TRAC-1BF and 100TRAC-1BF, the fracture energy was improved by 91% and 56% and the fracture toughness was enhanced by 69% and 43%, respectively. Furthermore, the added dosages of BF were found useful in improving the flexural strength, elastic modulus, and toughness indices of both types of recycled concretes. From the DIC analysis, it was found that the BF modified concrete contained continuous spread of strain contours throughout the beam depth and revealed higher strain values than the reference sample. Lastly, the microstructure observation demonstrated better concrete matrix quality and BF-cement matrix interaction which thereby confirms the usefulness of the applied strength enhancing technique for the application of high strength fiber reinforced recycled concrete.