Tri-dimensional (3D) cell aggregates or spheroids are considered to be closer to physiological conditions than traditional 2D cell culture. Mesenchymal stem cells (MSCs) assembling in spheroids have increased the survival of transplanted cells. The regulation of the biological processes that maintain crucial physiological reactions of MSCs is closely related to the functioning of ion channels. The pattern of expression, role and regulatory mechanisms of ion channels could be significantly different in 3D compared to 2D culture, and, thus, needed to be properly analyzed on the level of ionic currents. We developed a specific approach that allowed us to register, for the first time, endogenous ion channels in endometrial MSCs (eMSCs) assembled in spheroids. Particularly, using the single-channel patch-clamp technique, we have recorded the activity of ion channels and observed their functional interplay in mechanosensitive clusters. Our experimental protocol could be applied for identification and studying of ion channels in 3D cell cultures.

Recent improvements within the fields of high-throughput screening and 3D tissue culture have provided the possibility of developing in vitro micro-tissue models that can be used to study diseases and screen potential new therapies. This paper reports a proof of concept study on the use of microvalve-based bioprinting to create laminar MSC-chondrocyte co-cultures as an in vitro model of autologous chondrocyte implantation (ACI), an established cellular therapy for osteoarthritis. Microvalve-based bioprinting uses microvalves to deposit cells suspended in a liquid in a consistent and repeatable manner. In this case MSCs and chondrocytes have been sequentially deposited into an insert based transwell system in order to create a laminar co-culture, with variations in the ratios of the cell types used to investigate the potential for MSCs to stimulate improved repair. Histological and indirect immunofluorescence staining revealed the formation of dense tissue structures within the chondrocyte and MSC-chondrocyte cell co-cultures, alongside the establishment of a proliferative region at the base of the tissue. No stimulatory or inhibitory effect in terms of ECM production was observed through the introduction of MSCs, although the potential for an immunomodulatory benefit remains. This proof-of-concept study therefore provides a novel method to enable the scalable production of therapeutically relevant micro-tissue models that can be used for in vitro research to optimise ACI procedures.

The Atlantic cod (Gadus morhua) and red king crab (Paralithodes camtschaticus) processing wastes are massive and unutilized in the Murmansk region of Russia. The samples of skin-containing waste of Atlantic cod fillets production were hydrolyzed using enzyme preparations derived from red king crab hepatopancreases, porcine pancreases and Bacillus subtilis bacteria. The activity of enzymes from crab hepatopancreases was significantly higher than the activity of enzymes derived from other sources. The optimal conditions of the hydrolysis process have been figured out. The samples of cod processing waste hydrolysate were analyzed for amino acid composition and molecular weight distribution. The samples of hydrolysate were used as core components for bacterial culture medium samples. The efficiency of the medium samples was tested for Escherichia coli growth rate; the most efficient sample had efficiency 95.3% of that of a commercially available medium based on fish meal. Substitution of medium components with those derived from industrial by-products is one of the ways to decrease a cost of a culture medium in biopharmaceutical drug production.

Multi-step evolution characterizes the carcinogenesis of GI malignancies, where the disease progresses through the accrual of genetic mutations, co-option of multiple host cellular factors and the acquisition of increasingly aggressive pathological features. Ensuing cancer complexity and plasticity are challenges for therapeutic approaches. On the other hand, disease inception is often triggered by a defined oncogenic event, which leads to escalating effects on gene expression, cellular metabolism, chromatin organization and ultimately cell identity. Decoding this molecular cascade of events is crucial to design strategies for early diagnosis as well as to tackle tumor onset. To this aim, the in vitro culture of non-transformed epithelial cells represents a major technical challenge. Genetic studies, combined with lineage-tracing experiments demonstrated that pancreatic cancerous lesions originate from acinar cells, a specialized cell type in the pancreatic epithelium, following mutations of the KRAS oncogene. Ex vivo culture of pancreatic acinar cells has allowed the identification of early mutant KRAS-driven alterations. Primary acinar cells survive in vitro in organoid-like 3D spheroids, which can recapitulate histological and molecular features of disease initiation. Here, we will discuss the isolation and culture of primary mouse pancreatic acinar cells, providing and historical and technical perspective. Impact for pancreatic cancer research will also be debated; in particular we will dissect the role of KRAS signaling in driving oncogenic transformation and highlight differences with canonical 2D culture of established cancer cell lines.

With ongoing colony losses driven in part by the Varroa mite and the associated exacerbation of virus load, there is an urgent need to protect honey bees (Apis mellifera) from fatal levels of virus infection and from nontarget effects of insecticides used in agricultural settings. A continuously replicating cell line derived from the honey bee would provide a valuable tool for study of molecular mechanisms of virus – host interaction, for screening of antiviral agents for potential use within the hive, and for assessment of the risk of current and candidate insecticides to the honey bee. However, the establishment of a continuously replicating, honey bee cell line has proved challenging. Here we provide an overview of attempts to establish primary and continuously replicating hymenopteran cell lines, methods (including recent results) for establishing honey bee cell lines, challenges associated with the presence of latent viruses (especially Deformed wing virus), in established cell lines and methods to establish virus-free cell lines. We also describe the potential use of honey bee cell lines in conjunction with infectious clones of honey bee viruses for examination of fundamental virology.

Crotoxin (CTX), the predominant toxin in Crotalus durissus terrificus snake venom (CdtV), has anti-inflammatory and immunomodulatory effects. Despite its inhibitory action on neutrophil migration and phagocytosis, CTX does not directly affect the production of reactive oxygen species (ROS) by the neutrophils. In contrast, it enhances the generation of reactive oxygen and nitrogen intermediates by macrophages. Given the importance of macrophage-neutrophil interactions in innate antimicrobial defense, the aim of this study was to investigate the effect of CTX on neutrophil ROS production and killing activity, either through CTX-treated macrophage co-culture or conditioned medium of CTX-treated macrophages. The results showed an important modulatory action of CTX on the neutrophil function as well as neutrophil-macrophage interactions, as demonstrated by the increased production of hydrogen peroxide, hypochlorous acid, nitric oxide and TNF-α, along with the increased fungicidal activity of neutrophils.

The viability, productivity and survival of higher plants under the adverse factors influence are largely determined by the functional activity of the antioxidant system. The aim of our work was to investigate changes in formation of high-molecular (superoxide dismutase and peroxidase) and low-molecular (phenolics, including flavanols and proanthocyanidins) antioxidants in callus culture of Camellia sinensis under influence of phenolic precursors (L-phenylalanine – 3 mM, trans-cinnamic acid – 1 mM, naringenin – 0.5 mM). According to the data obtained, precursors effect on tea callus cultures did not lead to significant increasing of superoxide dismutase and peroxidase activity in most cases. However, it led to the increased accumulation of the total phenolics content, as well as flavanols and proanthocyanidins contents. For C. sinensis callus cultures, the most promising regulator of phenolic compounds was L-phenylalanine, in the presence of which its content increased almost twice. Thus, the exogenous effect of various precursors is possible to use for the targeted regulation of the certain phenolics classes accumulation in plant cells.

Plant biotechnology is dependent today on accessing broader plant genetic resources for food and agriculture (PGRFA) diversity for ensuring food security for our future. Heterogenous agricultural lands are considered today as hotspot for biodiversity, and they also exist in the hilly mountain areas of Romanian’s Carpathians. The scope of this review is to describe traditional home-hardens related to the land use of households and cultivated species from Moșna commune, Sibiu County, Romania. The survey was realized with the support of authorities and local stakeholders from 2010 up to 2019. The results of this survey revealed that maintenance of crop species structure and land use of traditional households is longer than 100 years. Consequently at least 15 traditional Saxon home-gardens of 120 taken into the study may be defined. Today these home-garden covers a media of 26% of the total area of a household and should be part of a surface ranging between 5,000 and 1,500 m2. The first Local Council Decision in Romania in 2019 supporting the recognition of landraces and home-gardens as heritage values was officially adopted supporting the need to more access bottom-up approaches for supporting biodiversity conservation.

This paper aims to stress the nexus between sustainable development, economic freedom and local participatory democracy in 21 European smart cities. The development of smart cities is strongly correlated with technological advancements and improved access to pub-lic services for all residents. Furthermore, smart cities are characterized by efficient man-agement of social, economic and environmental resources and a strong partnership be-tween the public and private sectors. The paper aims to map the dynamics of local partic-ipatory democracy in European smart cities and to identify several relevant predictors of local participatory democracy. The sample is represented by 21 European capitals (20 EU countries plus UK), which are included in the Smart City Index (SCI). Using the quantita-tive design based on both statistical regressions and structural equations modelling (SEM), the paper shows positive statistical correlation between economic factors, sustainable de-velopment, political culture, education and the level of local participatory democracy. Be-sides the exogenous variables, the study emphasized the main priority areas, which might be seen as urgent issues for local governments. The conclusion of the paper underlines the fact that economic freedom, sustainable development, upper secondary education, afford-able housing and an increased level of citizens’ engagement could predict further evolu-tions of the local participatory democracy in 21 European capitals.

Electrospinning is a simple versatile method to generate nanofibres. Remarkable progress has been made in the development of electrospinning process. The production of nanofibers is affected by many parameters, which influence the final material properties. Electrospun fibers have a wide range of applications such as energy storage and biomedical scaffolds. Polyacrylonitrile (PAN) has been widely used in the production of carbon nanofibers. PAN has received increasing interest in recent years, due to its excellent characteristics, such as spinnability, biocompatibility, and commercial viability, opening new applications in the biotechnological field. This paper provides an overview of the electrospinning process, its main parameters, and operation modes that affect nanofibers fabrication. A review of their biological applications is proposed. A focus is made on PAN fibers formation, functionalization, and application as scaffolds to allow cell growth. Overall, nanofibre scaffolds, made of stabilized PAN, appear as a powerful tool in medical applications needing controlled cell culture.

Since the 21st century, China's urbanization process has been rapid development, the concept and function of urban public space in the city has been gradually paid attention to. In order to guarantee life and water, most urban construction relies on rivers, and Bridges are the most important way to communicate between urban areas. The main functional part of the bridge is the span structure, that is, the bearing structure of the bridge, and the lower part of the "gray" space formed by the bridge structure. Considering the social level, with the economic growth and urbanization development, people have brought a better living environment and quality of life, and also improved the requirements for urban public environment. In the increasingly tense urban space, how to use and transform the space under the bridge is a problem that needs to be considered and solved. In view of this problem, in this study, we try to solve the micro-transformation of space under Bridges in cities through public art from the perspective of "regional culture" and "art for all". This paper analyzes the micro-transformation of space art under Bridges in two large cities of Shanghai and Foshan, namely, the space under Bridges under Songhong Road in Shanghai, the space under Bridges under Central Of Suzhou River and the space under Bridges under Pingsheng Bridge in Foshan. This paper discusses the cultural intervention of "regional culture" in the micro-transformation of the space under the bridge, and the influence and effect of "art for all" on the public art space under the bridge after the transformation to the community and the public.

Short chain quinones (SCQs) have been identified as potential drug candidates against mitochondrial dysfunction, which is largely dependent on their reversible redox characteristics of the active quinone core. We recently synthesized a SCQ library of > 148 naphthoquinone derivatives and identified 16 compounds with enhanced cytoprotection compared to the clinically used benzoquinone idebenone. One of the major drawbacks of idebenone is its high metabolic conversion in the liver, which significantly restricts is therapeutic activity. Therefore, this study assessed the metabolic stability of the 16 identified naphthoquinone derivatives 1-16 using hepatocarcinoma cells in combination with an optimized reverse-phase liquid chromatography (RP-LC) method. Most of the derivatives showed significantly better stability than idebenone over 6 hours (p < 0.001). By extending the side-chain of SCQs, increased stability for some compounds was observed. Metabolic conversion from the derivative 3 to 5 and reduced idebenone metabolism in the presence of 5 were also observed. These results highlight the therapeutic potential of naphthoquinone-based SCQs and provide essential insights for future drug design, prodrug therapy and polytherapy, respectively.

Cancer cell lines are widely used as in vitro models of tumorigenesis, facilitating fundamental discoveries in cancer biology and translational medicine. Currently, there are few options for glioblastoma (GBM) treatment and limited in vitro models with accurate genomic and transcriptomic characterization. Here, a detailed characterization of a new GBM cell line, namely AHOL1, was conducted in order to fully characterize its molecular composition based on its copy number alteration (CNA) and transcriptome profiling, followed by the validation of key elements associated with GBM tumorigenesis. Large numbers of CNAs and differentially expressed genes (DEGs) were identified. CNAs were distributed throughout the genome, including gains at Xq11.1-q28, Xp22.33-p11.1, Xq21.1-q21.33, 4p15.1-p14, 8q23.2-q23.3 and losses at Yq11.21-q12, Yp11.31-p11.2 and 12p13.31 positions. Nine druggable genes were identified, including HCRTR2, ETV1, PTPRD, PRKX, STS, RPS6KA6, ZFY, USP9Y and KDM5D. By integrating DEGs and CNAs, we identified 57 overlapping genes enriched in fourteen pathways. Altered expression of several cancer-related candidates found in the DEGs-CNA dataset was confirmed by RT-qPCR. Taken together, this first comprehensive genomic and transcriptomic landscape of AHOL1 provides unique resources for further studies and identifies several druggable targets that may be useful for therapeutics and biologic and molecular investigation of GBM.

This is a theoretical and empirical article. In the first part, an original definition of the cross-border market for cultural services has been developed, based on classic market definitions. The structure and conditions of functioning for this market have also been presented. The next part of the article discusses the supply side of the cross-border market of cultural services for the two twin towns: Cieszyn (Poland) and Český Těšín (Czech Republic). Next, based on the surveys conducted, the participation of residents of both towns in the most important cultural events organised by cultural institutions on both sides of the border was examined.

Background: Metabolic flux analysis (MFA) is an indispensable tool in metabolic engineering. The simplest variant of MFA relies on an overdetermined stoichiometric model of the cell’s metabolism under the pseudo-steady state assumption, to evaluate the intracellular flux distribution. Despite its long history, the issue of model error in the overdetermined MFA, particularly misspecifications of the stoichiometric matrix, has not received much attention. Method: We evaluated the performance of statistical tests from linear least square regressions, namely Ramsey RESET test, F-test and Lagrange multiplier test, in detecting model misspecifications in the overdetermined MFA, particularly missing reactions. We further proposed an iterative procedure using the F-test to correct such an issue. Result: Using Chinese hamster ovary and random metabolic networks, we demonstrated that: (1) a statistically significant regression does not guarantee high accuracy of the flux estimates, (2) the removal of a reaction with a low flux magnitude can cause disproportionately large biases in the flux estimates, (3) the F-test could efficiently detect missing reactions, and (4) the proposed iterative procedure could robustly resolve the omission of reactions. Conclusion: Our work demonstrated that statistical analysis and tests could be used to systematically assess, detect and resolve model misspecifications in the overdetermined MFA.

Salidroside, a valuable phenylethanoid glycoside, is obtained from plants belonging to the Rhodiola genus, known for its diverse biological properties. At present, salidroside is still far from large-scale industrial production due to the lower titer and higher process cost. In this study, we constructed a biosynthetic pathway from L-tyrosine to salidroside in Saccharomyces cerevisiae. To increase sali-droside production, we for the first time constructed an in vivo UDP-glucose regeneration system that sucrose synthase GmSUS worked in conjunction with UDP-glucose transferase RrUGT33 to improve the UDP-glucose availability with inexpensive sucrose as the consumed substrate. Finally, the molar conversion rate of L-tyrosine could reach 95%. To further reduce the cost, co-culture with E. coli that synthesized L-tyrosine de novo resulted in an 81% reduction in the final substrate cost per 1 kg of salidroside compared to the initial cost. The final titer of salidroside obtained 3.8 g/L and was the highest titer achieved at the Shake-Flask-Level in the existing reports. And this marked the first successful synthesis of salidroside in an in situ enhanced UDP-glucose system using sucrose. This accomplishment laid a robust groundwork for further investigations into the synthesis of other notable glycosides and natural compounds.

This study, which was supported by a quantitative research paradigm with two experiment and two control groups of 4x4 quasi-experimental design, aimed to find out the impact of a sustainable progressive STEAM (SP-STEAM Model) application model on 5th grade primary school students critical thinking dispositions and mathematic achievements in Northern Cyprus. The treatment model was applied to experimental groups for 14 weeks of time. Split-plot multiple group analysis of variance (Split-plot ANOVA) statistical technique was used to calculate between and within group significances regarding exogenous variable. SPSS24 software package was used for the analysis. Pre-test and post-test results derived from experiment and control groups revealed significant effect of SP-STEAM Model on 5th grade primary school students’ critical thinking dispositions as measured with CCTDI and mathematic achievements as measured with a dedicated ex-am. The results were discussed in detail under the light of related literature and suggestions for further studies were proposed.

This article offers a theoretical framework to contextualise the concept of "Safety Culture" in the field of public safety and Disaster Risk Reduction (DRR), with the aim of supporting its understanding and measurement in our communities and finally attaining more disaster resilient societies. The work discusses the role of cultural dimensions in risk management and DRR and highlights the relevance of building knowledge and practices based on the consideration of culture and cultural variability in all phases of the risk management process. After an overview of the Safety Culture concept, including its origins, developments and applications, the text explores the transferability of this concept to the context of public safety. Then, based on the existing Safety Culture models, metrics and indicators from other sectors, the paper proposes an original definition of Safety Culture for the DRR context, shaped on a series of elements and dimensions specifically outlined for this context. Finally, the expected benefits of the application of Safety Culture concepts and techniques for further advances in DRR is discussed.

Since 2000, caffeine is used as citrate salt of caffeine for treatment of apnea of prematurity. However, it remained elusive if caffeine citrate has a direct function in the respiratory epithelium, potentially as modulator for ciliary beat frequency (CBF) regulation in human respiratory cells (hRECs). Here, we studied the mechanism of action of caffeine citrate on the respiratory epithelium on hRECs derived from healthy donors and individuals with cystic fibrosis (CF) cultured under air-liquid-interface (ALI) conditions. CBF analyses of hRECs were performed by high-speed video microscopy. In the presence of caffeine, hRECs show a significant increase in CBF over ciliary base frequency (CBFb), demonstrating a direct influence of caffeine on ciliary function. We also demonstrate a specific action of caffeine at ryanodine receptor (RYR) channels, mediating calcium efflux from intracellular stores to increase CBF. We describe a novel pharmacological effect of caffeine citrate - direct increase of CBF of primary hRECs cultured at ALI. Caffeine citrate has certain advantages over other methylxanthines: it was shown to have stable plasma concentration, longer half-life, and is better tolerated than e.g. theophylline. We conclude that caffeine citrate therapy might be suitable strategy to treat respiratory diseases with compromised mucociliary clearance.

Marine invertebrates are model organisms in several areas of biological sciences, being a source of massive biological information. Although, the scientific relevance of marine invertebrates, the research with them can be limited for their tissue characteristics and troubles for the replication of physical and chemical properties of seawater. Thence, the main goal of this laboratory workflow is to provide a useful methodological approach to reduce the experimental limitations during the study of marine invertebrates. The present study describes experimental methodologies for the collection, transport, and maintenance of sessile tunicates. Also, an approach to observe and characterize, a diverse population of blood cells in marine invertebrates, by several cytological stains and electron microscopy. Lastly, suggestions and protocols to extract quality RNA from samples with high concentrations of salts, pigments, secondary metabolites, and polysaccharides. This methodological approach can be easily adapted to other marine invertebrates, moreover uses low-cost reagents and widely available laboratory equipment. Making possible the study of different types of marine animals in diverse locations.

The tumor organoid (tumoroid) model in three-dimensional (3D) culture systems has been developed to reflect more closely the in vivo tumors than 2D-cultured tumor cells. Notably, extracellular vesicles (EVs) are efficiently collectible from the culture supernatant of gel-free tumoroids. Matrix metalloproteinase (MMP) 3 is a multi-functional factor playing crucial roles in tumor progression. However, roles of MMP3 within tumor growth and EVs have not unveiled. Here, we investigated the protumorigenic roles of MMP3 on integrities of tumoroids and EVs. We generated MMP3-knockout (KO) cells using the CRISPR/Cas9 system from rapidly metastatic LuM1 tumor cells. Moreover, we established fluorescent cell lines with palmitoylation signal-fused fluorescent proteins (tdTomato and enhanced GFP). Then we confirmed the exchange of EVs between cellular populations and tumoroids. LuM1-tumoroids released large EVs (300-1000 nm) and small EVs (50-200 nm) while the knockout of MMP3 resulted in the additional release of broken EVs from tumoroids. The loss of MMP3 leads to a significant reduction in tumoroid size and to the development of the necrotic area within tumoroids. MMP3 and CD9 (a category-1 EV marker tetraspanin protein) were significantly down-regulated in MMP3-KO cells and their EV fraction. These weakened phenotypes by MMP3 KO were markedly rescued by the addition of MMP3-rich EVs or conditioned medium (CM) collected from LuM1-tumoroids, which caused a dramatic rise in the expression of MMP3, CD9, and Ki-67 (a marker of proliferating cells) in the MMP3-null/CD9-low tumoroids. Notably, MMP3 enriched in tumoroids-derived EVs and CM deeply penetrated into recipient MMP3-KO tumoroids, resulting in a remarkable enlargement of solid tumoroids, while MMP3-null EVs did not. These data demonstrate that EVs can mediate molecular transfer of MMP3 resulting in increasing the proliferation and CD9+ tumorigenesis, indicating crucial roles of MMP3 in tumor progression.

Within the framework of theoretical developments in so-called participatory culture and the context of funds of identity, incorporated within what is known as the funds of knowledge approach, an innovative teaching methodology is implemented that allows students to actively participate in knowledge construction. The project translates into a proposal for educational contextualization and personalization based on the students’ funds of identity, that is, those resources (people, artifacts, places, activities, institutions) they consider to be most relevant and significant to define themselves. Once these have been identified through identity artifacts, such as collages or self-drawings, students link some of these funds of identity to curricular content of the subject and produce a video that shows the results of this academic work. The final product is shared on a YouTube channel containing the videos of all of the students in the class. The phases of the project are described and illustrated. We argue that the proposed teaching and learning project, which is cross-disciplinary in nature, allows for principles such as educational contextualization, funds of identity, and participatory culture to be incorporated into what we call here the funds of identity 2.0 approach, putting it into educational practice.

This paper examines cognitive biases which affect the ability of decision makers to make rational decisions in an organizational context. The motivation for this analysis begins with the observation of catastrophic accidents caused by human error but in an organizational context. This paper expands on the concept of cognitive bias to define organizational biases which are the factors that affect decisions in an organizational context. The paper distinguishes between organizational biases, which are the focus of this paper, and individual biases, which are biases experienced by individuals but may have organizational consequences. The purpose of this paper is to identify methods to mitigate the risks of organizational accidents, accidents which involve many people operating at different levels of an organization. The methodology is to identify those decisions that would address the specific organizational biases. The focus of this paper is the decisions for mitigating the risks associated with decisions in an organizational context. Results are shown for seven organizational biases, six specific case studies, and four decision options. This paper concludes that organizational biases are intrinsically different from individual biases and that these differences lead to different decision options from those that mitigate individual biases; however, they may exist concurrently.

Endophytic fungi colonize the inner tissues of plants and provide direct and indirect benefits to the plant. Although Mongolia is rich in medicinal plants, due to climatic and anthropogenic reasons the resources are being depleted and many species are under the threat of gradual extinction, while the endophytic fungi of Mongolian plants are largely unknown. Understanding the diversity and metabolic potential of endophytic fungi is of great significance in terms of the utilization of beneficial fungal strains in plant propagation and the discovery of bioactive compounds. In this study, a total of 24 culturable endophytic fungal strains were isolated from Cynomorium songaricum (Rupr.), a medicinal and vulnerable plant species of Mongolia. Based on the morphological characteristics and the sequences of the rDNA internal transcribed spacer (ITS) region, the isolates were identified to 6 genera: Fusarium (8), Clonostachys (7), Penicillium (6), Alternaria (1), Aspergillus (1), and Madurella (1). The antimicrobial activity was assessed by agar-diffusion method, revealing that 15 strains were able to inhibit the growth of at least one of the test organisms. Among them, 1 strain showed inhibitory activity against Escherichia coli, 12 against Bacillus subtilis, 13 against Staphylococcus aureus, 8 against Aspergillus niger, respectively. All fungal strains were screened for their ability to solubilize complex phosphorus and zinc minerals and for their production of indole-3-acetic acid (IAA). The strains were preserved in the national culture collection for further sustainable exploitation of their biotechnological potential.

This paper examines the learning experience of an inter-faculty higher education course conducted in an expanded remote format, using digital space as facilitator. The study focuses on the implementation of personalized education, a pedagogical approach that emphasizes the diversification of resources, strategies, and actions to empower learners in assigning personal value and meaning to their learning journey. By presenting an action research project, this study highlights the pivotal role of personalized education in guiding the course design. To foster an engaging learning environment, the course incorporated open activities and leveraged tools and platforms of the social web. Through these means, the participants, forming a community of practice, were able to collaboratively generate knowledge before, during, and after synchronous sessions. Furthermore, the participants effectively disseminated this knowledge through networks and engaged in discussions with external stakeholders. The study underscores how personalized education facilitates situated and meaningful learning by fostering expanded, flexible, and distributed experiences. These experiences revolve around a transmedia narrative that seamlessly integrates digital culture into learning opportunities and processes, effectively naturalizing its presence. The results highlight the value of personalized education approaches in adapting to the complexities of the digital age and establishing meaningful learning experiences for students.

The cell concentration measurement on a culture assay using bioimpedance is a very useful tool, but it is complex to translate impedance to cell concentration values. The purpose of this work is to find a method to obtain in real time the cell concentration values for a given cell-culture assay using an oscillator as the measurement circuit. From a basic cell-electrode model, enhanced models of the cell culture immersed in a saline solution (culture medium) can be derived. These models can be used in a fitting routine to real time estimation of the cell concentration in a cell culture, using the oscillation frequency and amplitude delivered by measurement circuits proposed by the authors. Based on real experimental data (frequency and amplitude of oscillations) obtained by connecting the cell culture to an oscillator as a load, the fitting routine is simulated, and real time data of cell concentration is achieved. These results are compared with concentration data found by traditional optical methods for counting. In addition, the error obtained is divided and analyzed in two parts: in the first part of the experiment (when the few cells are adapting to the culture medium) and the second part of the experiment (when the cells grow exponentially until they completely cover the well).Low error values are obtained in the growth phase of the cell culture (the relevant phase), therefore the results obtained are considered to be promising, proving that the fitting routine is valid, and that the cell concentration can be measured in real time using an oscillator.

This study focused on the characterization of lactic acid bacteria (LAB) in a renowned traditional Italian cheese, Pecorino di Farindola, to select an autochthonous culture and investigate its po-tential for the improvement of safety and functional properties. Two hundred and six LAB isolated throughout production and maturation from nine cheese lots of three farms were identified by 16S rRNA gene sequencing and tested for the presence of genes encoding virulence factors, vancomycin resistance (for enterococci), biogenic amines (BAs) and bacteriocin production, for antimicrobial activity, and for the capacity to survive in the gastro-intestinal tract (GIT) based on tolerance to low pH and bile salts and adhesion to CaCo-2 cells. A Lacticaseibacillus paracasei isolate was used in cheese making and determined a decline of spiked Listeria monocytogenes and Escherichia coli O157 faster than in control cheese. The autochthonous bacterial groups were numerically unaffected, apart from lactobacilli that were recovered in higher numbers in the cheese with added L. paracasei. Based on Repetitive Extragenic Palyndrome (Rep) PCR profiles, the added strain possibly dominated until day 88 in cheese. Results encourage further trials with autochthonous cultures to efficiently inhibit hazardous bacteria and enrich a functional microbiota in Pecorino di Farindola and similar cheeses.

(1) Background: Nursing homes’ preparedness in managing a public health crisis has been fragile, with effects on safety culture. The objective of this study was to assess nursing homes’ COVID-19 preparedness in Southern Portugal, including personnel’s work experiences during the pandemic. (2) Methods: We used a COVID-19 preparedness checklist to be completed by management teams, followed by follow-up calls to nursing homes. Thereafter, a survey to personnel was applied. Data analysis included descriptive statistics, exploratory factor analysis, and thematic analysis of open-end questions. (3) Results: In total, 71% (138/195) of eligible nursing homes returned the preparedness checklist. We conducted 83 follow-up calls and received 720 replies to the personnel survey. On average, 25% of nursing homes did not have an adequate decision-making structure to respond to the pandemic. Outbreak capacity and training were fragile areas among nursing homes’ contingency plans. We identified compliance with procedures and nonpunitive response to mistakes as fragile areas of safety culture, and teamwork as a strong safety area. (4) Conclusions: To strengthen how nursing homes cope with upcoming phases of the COVID-19 pandemic or future public health emergencies, nursing homes’ preparedness and safety culture should be fostered and closely monitored.

Many individuals are wearing face masks improperly at ‘half mask’ and potentially breathing out microbes that can contaminate the air as well as anything below the nose. This note provides the first report that bacteria and fungi breathed out during nasal air exhalation are able to be cultured after landing on blood agar plates. The CFU’s are higher after both 10 breaths and extremely significant for 20 breaths compared to the control plates exposed to the air. Implications of this finding are that going ‘nose commando’ may be able to continue the spread of respiratory diseases such as COVID-19.

This article presents the study of religious pluralism and decolonization of Indian mind in Russia. The paper analyzes the investigation results concerning psychological features of modern Indian students from universities in Russia and India. For measuring of connection between religious pluralism and decolonization of Indian mind we made socio-psychological investigation of Indian students. We made 254 semi-structured interview with Indian students who are studying in Russia and India. According to the result of investigation that decolonization of Indian mind is connected with the level of religious pluralism. Among the values principles of religious pluralism get more significance and importance in decolonizing mind of Indian students.

The “stem cells” discipline represents one of the most dynamic areas in biomedicine. While adult marine/aquatic invertebrate stem cell (MISC) biology is of prime research and medical interest, studies on stem cells from organisms outside the classical vertebrate (e.g., human, mouse, zebrafish) and invertebrate (e.g., Drosophila, Caenorhabditis) models have not been pursued vigorously. Marine/aquatic invertebrates constitute the largest biodiversity and the widest phylogenetic radiation on Earth, from morphologically simple organisms (e.g. sponges, cnidarians), to the more complex mollusks, crustaceans, echinoderms and protochordates. These organisms illustrate a kaleidoscope of MISC-types that participate in the production of a large number of novel bioactive-molecules, many of which are of significant potential interest for human health. MISCs further participate in aging and regeneration phenomena, including whole-body regeneration. For years, the European MISC-community has been highly fragmented and scarce ties were established with biomedical industries in attempts to harness MISCs for human welfare. Thus, it is important to: i) consolidate the fragmented European community working on MISCs; ii) promote and coordinate European research on MISC biology; iii) stimulate young researchers to embark on research in MISC-biology; iv) develop, validate, and network novel MISC tools and methodologies; v) establish the MISC discipline as a forefront interest of biomedical disciplines, including nanobiomedicine; vi) establish collaborations with industries to exploit MISCs as sources of bioactive molecules. In order to fill the recognised gaps, the EC-COST Action 16203 “MARISTEM”, has recently been launched. At its initial stage the consortium unites 26 scientists from EC countries, Cooperating countries and Near Neighbor Countries.

Background. Transferrin receptor 1 (TfR1), glucose transporter 1 (GLUT1), and CD98hc are candidates for targeted therapy at the blood-brain barrier (BBB). Our objective was to challenge the expression of TfR1, GLUT1, and CD98hc in brain capillaries by the histone deacetylase inhibitor (HDACi) valproic acid (VPA). Methods. Primary mouse brain capillary endothelial cells (BCECs) and brain capillaries isolated from mice injected intraperitoneally with VPA were examined using RT-qPCR and ELISA. Targeting to the BBB was performed by injecting monoclonal anti-TfR1 (Ri7217)-conjugated gold nanoparticles measured using ICP-MS. Results. In BCECs co-cultured with glial cells, Tfrc mRNA expression was significantly higher after 6 h VPA returning to baseline after 24 h. In vivo Glut1 mRNA expression was significantly higher in males, but not females', receiving VPA, whereas Cd98hc mRNA expression was unaffected by VPA. TfR1 increased significantly in vivo after VPA, whereas GLUT1 and CD98hc were unchanged. Uptake of anti-TfR1-conjugated nanoparticles was unaltered by VPA in spite upregulated TfR expression. Conclusions. VPA upregulates TfR1 in brain endothelium in vivo and in vitro. VPA does not increase GLUT1 and CD98hc proteins. The increase in TfR1 does not result in higher anti-TfR1 antibody targetability, suggesting targeting sufficiently occurs with available transferrin receptors without further contribution from accessory VPA-induced TfR1.

This article explores the concept of sustainable leadership in the digital era and the role of technology in promoting environmental sustainability. As the world becomes increasingly interconnected through digital advancements, leaders are facing new challenges and opportunities to address environmental concerns. This article examines the key characteristics of sustainable leadership and discusses how technology can be utilized to promote environmental sustainability. It highlights various technological innovations and strategies that leaders can employ to foster a sustainable organizational culture. Moreover, the article discusses potential barriers and limitations of technology in achieving sustainability goals. Overall, it emphasizes the significance of sustainable leadership and technology integration in addressing environmental challenges in the digital era.

Objectives: The study aimed to uncover the percentages of battered women in Jordan, its causes, forms, and relationship to several social factors. Methodology: The study was conducted on a random, targeted sample of (1308) women based on social sample survey method and electronic questionnaire tool for data collection, the Descriptive statistical method and chi-square test were used to examine statistically significant differences. Results: The study found that the percentage of battered women reached 17.1% during Corona pandemic in 2020, and the increase in men's violence against women during this period of was in large and medium degrees according to the sample. Husbands were the most practicing violence against their wives at 37.5%, followed by fathers against daughters at 28.6%, and brothers against sisters at 26.8%. The causes of male violence are due to social factors represented by male domination culture, interference by family and relatives, and economic factors represented by the high cost of living, low household income and poverty. The most common forms of violence are verbal, physical, and psychological. The study found that women in southern Jordan, who are poorer and less educated, and who live in a large family, are the most abused. The study recommended activating family and community service and reform offices, solving problems of deteriorating economic situation and individual poverty, and enlightening community awareness of the dangers of domestic violence.

The aim of this study was to investigate the role of DNA methylation in the regulation of in vitro and in vivo cartilage formation. Based on the data of an RNA chip-assay performed on chondrifying BMP2-overexpressing C3H10T1/2 cells, the relative expression of Tet1 (tet methylcytosine dioxygenase 1), Dnmt3a (DNA methyltransferase 3) and Ogt (O-linked N-acetylglucosamine transferase) genes was examined with RT-qPCR in mouse cell-line based and primary micromass cultures. RNA probes for in situ hybridization were used on frozen sections of 15-day-old mouse embryos. DNA methylation was inhibited with 5-azacytidine during culturing. We found very strong but gradually decreasing expression of Tet1 throughout the entire course of in vitro cartilage differentiation along with strong signals in the cartilaginous embryonic skeleton. Dnmt3a and Ogt expressions did not show significant changes with RT-qPCR and gave weak in situ hybridization signals. Inhibition of DNA methylation applied during early stages of differentiation reduced cartilage-specific gene expression and cartilage formation. In contrast, it had stimulatory effect when added to differentiated chondrocytes. Our results indicate that the DNA demethylation-inducing Tet1 is a significant epigenetic factor of chondrogenesis, and inhibition of DNA methylation exerts distinct effects in different phases of in vitro cartilage formation.

Thai basil is a renowned medicinal plant and a rich source of bioactive antioxidant compounds having several health benefits, with actions to prevent of cancer, diabetes and cardiovascular disease. Plant cell and tissue culture technologies can be routinely established as an important, sustainable and low-cost biomass source for the production of high-value phytochemicals. The current study aimed at developing an effective protocol for the production of Thai basil leaf derived callus cultures with sustainable and high production of biomass and antioxidants as an alternative of leaves production. MS basal medium with various concentrations of plant growth regulators (PGRs) compatible with nutraceutical applications (i.e., gibberellic acid (GA3) and 6-benzylaminopurine (BAP) either alone or in combination with naphthalene acetic acid (NAA)) were evaluated. Among all tested PGRs, the combination BAP:NAA (5 mg/L:1 mg/L) yield maximum biomass accumulation (fresh weight (FW): 190 g/L and dry weight (DW): 13.05 g/L) as well as enhanced phenolic (346.08 mg/L) production. HPLC quantification analysis indicated high productions of chicoric acid (35.77 mg/g DW) and rosmarinic acid (7.35 mg/g DW) under optimized callus culture conditions. Antioxidant potential was assessed using both in vitro cell free and in vivo cellular antioxidant assays. Maximum in vitro antioxidant activity DPPH (93.2 % of radical scavenging activity) and ABTS (1322 µM Trolox equivalent antioxidant capacity) was also observed for the extracts from callus cultures grown on optimal conditions. In vivo cellular antioxidant activity assay confirmed the effective protection against oxidative stress of the corresponding extract by the maximum inhibition of ROS and RNS production. Compared to commercial leaves, callus extracts showed higher production of chicoric acid and rosmarinic acid associated with higher antioxidant capacity. In addition, this biological system also has a large capacity for continuous biomass production, thus demonstrating its high potential for possible nutraceutical applications.

Breast cancer treatment mostly revolved around radiation therapy and surgical interventions, these treatments doesn’t provide satisfactory relief to the patients and carry unmanageable side-effects. Nanomaterials show promising results in treating cancer cells and have many advantages such as high biocompatibility, bioavailability and effective therapeutic capabilities. Interestingly, fluorescent magnetic nanoparticles have been used in many biological and diagnostic applications, but there is no report of use of fluorescent magnetic submicronic polymer nanoparticles (FMSP-nanoparticles) in the treatment of human breast cancer cells. In the present study, we have tested the effect FMSP-nanoparticles on human breast cancer cells (MCF-7). We have tested different concentrations (1.25µg/1mL, 12.5µg/mL and 50µg/1mL) of FMSP-nanoparticles in MCF-7 cells and evaluated the nanoparticles response morphometrically. Our results revealed that FMSP-nanoparticles produced a concentration dependent effect on the cancer cells, dose of 1.25µg/mL produced no significant effect on the cancer cell morphology and cell death, whereas dosages of 12.5µg/mL and 50µg/mL respectively showed significant nuclear augmentation, disintegration, chromatic condensation followed by dose dependent cell death. Our results demonstrate FMSP-nanoparticles have ability to induce cell death in MCF-7 cells and may be considered as a potential anti-cancer agent for breast cancer treatments.

The study was aimed at examining the status of research productivity and exploring the factors behind the identified status in Ethiopian universities from SDT and RBVT perspectives. In phase I, a null hypothesis was tested using a one-sample t-test at 0.05 alphas while in phase II one RQ was solved using qualitative analysis. The sample involved 531 academics, of whom 13 were interviewed. The mixed-methods research approach and explanatory sequential design guided this research. The status of research project production, indexed publication, and conference presentation performances in Ethiopian universities demonstrated statistically significant mean differences below the institutional, national, and world average cut-off points (performing at least one research project, one indexed journal, and one conference paper presentation per year per academic staff). Furthermore, we explored scarcity of funds, low research capacity, weak research culture, lack of productive institutional leadership, and political instability in the country as major causes of the low research productivity status. The researchers felt that the study would contribute to both promoting academic staff's research productivity and enriching the higher education research literature. MOE, university presidents, and deans should intervene in causes to improve academics’ research productivity status towards the desired local and global standards.

Researchers have developed and used several three-dimensional (3D) culture systems, including spheroids, organoids, and tumoroids. Drug resistance is a crucial issue involving recurrence in cancer patients. Many studies on anticancer drugs have been done in 2D culture systems, where-as 3D cultured tumoroids have many advantages for assessing drug sensitivity and resistance. Here, we aim to investigate whether Cisplatin (a DNA crosslinker), Imatinib (a multiple tyro-sine kinase inhibitor), and 5-Fluorouracil (5-FU: an antimetabolite) alter tumoroid growth of metastatic colorectal cancer (mCRC). To establish a liquid-based 3D multiplexing reporter assay system, LuM1 (a murine mCRC cell line) was stably transfected with the Mmp9 promoter-driven ZsGreen reporter gene, which was designated as LuM1/m9 cells and cultured in NanoCulture Plate (NCP), a 3D culture device. The larger tumoroids were not sensitive to Cisplatin and ex-pressed ABCG2 (a marker of cancer stem cells, a.k.a. a drug efflux transporter), whereas smaller cell-aggregates were more sensitive to Cisplatin. Both Imatinib and Cisplatin significantly in-creased tumoroid growth (larger than 300 μm2) and Mmp9 promoter activity and were not cytotoxic to the mCRC tumoroids. On the other hand, 5-FU was cytotoxic to the tumoroids and significantly inhibited tumoroid growth, although not completely. Thus, platinum resistance and imatinib resistance in mCRC were modeled using the liquid-based 3D cultured tumoroid system. The tumoroid culture is useful and easily accessible for the assessment of drug sensitivity and resistance.

In this digital world, organisations are facing global competition as well as manpower pressures leading towards knowledge economy, which heavily impacts on their local and international businesses. The trend is to foster collaboration and knowledge sharing to cope with these problems. With the advancement of technologies and social engineering that can connect people in virtual world across time and distance, several organisations are embarking on knowledge management (KM) systems, implementing community of practice (CoP) approach. However, virtual communities are relatively new paradigms, and there are several challenges to their successful implementation from an organisation’s point of interest. There is lack of CoP implementation framework that can cater to today’s dynamic business and sustainability requirements. To fill the gap in literature, this paper develops a practical framework for a CoP implementation with a view to align KM strategy with business strategy of an organization. It explores the different steps of building, sharing and using tacit and explicit knowledge in CoPs by applying Wiig KM cycle. It proposes a practical CoP implementation framework that adopts the Benefits-Tools-Organisation-People-Process (BTOPP) model in addressing the key questions surrounding each of the BTOPP elements with a structured approach. Finally, it identifies key challenges such as organizational culture and performance measurements, and provides practical recommendations to overcome them for a successful CoP implementation.

To proactively manage information security, enterprises often employ information security risk assessment techniques. Asset value - which is used to calculate the financial impact of possible threats - is one of the key parameters of information security risk. However, assets in a system are rarely independent, and their values are typically interdependent. Asset owners and IT teams may hold different views as regards these values, and there is thus the need to reduce subjectivity in a qualitative risk assessment. The research entails the development of a conceptual framework derived from the literature to minimize subjectivity, and the design of a system based on those concepts. The study uses the Unified Modeling Language as a design tool and puts forward an object-oriented model for defining asset values, in which the relationships between assets, vulnerabilities and related threats are identified. A “segregation of duties” approach is integrated into the risk management system to mitigate against subjectivity and better determine asset values. Survey responses from 16 practitioners working in the private and public sectors confirm the validity of the approach, but suggest it may be more workable in larger organisations where resources allow dedicated risk professionals to operate.

Truffles are ascomycete hypogeous fungi belonging to the Tuberaceae family of the Pezizales order that grow in ectomycorrhizal symbiosis with tree roots and are known for their peculiar aromas and flavors. Axenic culture of truffle mycelium is problematic because it is not possible in many cases, and the growth rate is meager when it is possible. This limitation prompts searching and characterizing new strains that can be handled in laboratory conditions for basic and applied studies. In this work, a new strain of Tuber borchii (strain SP1) has been isolated and cultured, and its transcriptome has been analyzed under different in vitro culture conditions. The results show that the best T. borchii SP1 growth was obtained using maltose-enriched cultures made with soft-agar and in static submerged cultures made at 22ºC. The transcriptome analysis of this strain cultured in different media indicated that most of the gene transcription effort is due to a limited number of genes (20% of genes account for 80% of the transcription), that the transcription profile of the central metabolism genes was similar in the different conditions analyzed with a transcription signal detected for around 80% of the annotated genes. The gene expression profile suggests that T. borchii uses a fermentative rather than respiratory metabolism, even in aerobic conditions. Finally, there is a reduced expression of genes belonging to secondary metabolite clusters, whereas there is a significative transcription of those involved in producing volatile aromatic compounds.

In this article, we have tried to answer the questions of how an entire city's fiction was created for the development of Japanese American internment camps and in which way contributions of sustainable crafts and even art could have interceded. To this aim, we have searched the literature and reviewed archives, primarily from the American West Coast. We have found that beyond adaptation to the circumstances, the visual representation of the city's settlement, founding, and daily activities, instead of typical panoptic or somber prisons’ imagery, are depicted in the selected images and that their use precisely fostered its sustainability. This leads us to deduce that a "city fiction" was necessary for such survival and endurance and that its artistic representation was primarily involved in the state's ideological apparatus. On the other hand, fissures subtly seethed the violence exerted in the camps by stealth and in a hidden fashion. We have concluded that artistic activity justified the city's fiction, together with other examples, highlighting the conditions of systemic violence and oppression for the inmates. In this framework, the artworks generated constitute a historical document of sustainability.

The concept of sustainable management in culture has been recognised in global strategic documents on sustainable development for more than a decade. It is also increasingly reflected in the cultural polices of particular states, and – very importantly –cultural managers responsible for shaping the cultural offer in cities are becoming more interested in this concept. Despite the increasing attention being paid to this topic among both practitioners and theoreticians of management, in none of these documents or other works can we find any content directly related to the possibility of applying this concept in a town which, due to political turmoil, has been divided by a national border. Hence, this gap was the direct impulse for taking up research in this field. In the article, by using different notions of the market, our own definition of a cross-border market for cultural services was developed, and the conditions for the functioning of this market were presented based on the example town of Cieszyn (Poland) and Český Těšín (Czech Republic). In the opinion of the authors of the article, the development and functioning of a cross-border market for cultural services is essential for the application of the concept of sustainable management of the cultural offer in a town divided by a border. For the purpose of the article, a survey and individual interviews with experts shaping the cultural offer in Cieszyn and Český Těšín were conducted. The results of the research prove that despite numerous cross-border Czech-Polish projects carried out by cultural institutions, there are still many barriers in the town which make it difficult for the residents to benefit from the cultural offer available on the other side of the border. These barriers limit the full implementation and application of the concept of sustainable management of the cultural offer.

Colonization of distant organs by tumor cells is a critical step of cancer progression. The initial avascular stage of this process (micrometastasis) remains almost inaccessible to study due to the lack of relevant experimental approaches. Here, we introduce an in vitro/in vivo model of organ-specific micrometastases of triple-negative breast cancer (TNBC) that is fully implemented in a cost-efficient chick embryo (CE) experimental platform. The model is built as three-dimensional (3D) tissue engineering constructs (TECs) combining human MDA-MB-231 cells and decellular-ized CE organ-specific scaffolds. TNBC cells colonized CE organ-specific scaffolds in 2-3 weeks, forming tissue-like structures. The feasibility of this methodology for basic cancer research, drug development and nanomedicine was demonstrated on a model of hepatic micrometastasis of TNBC. We revealed that MDA-MB-231 differentially colonize parenchymal and stromal com-partments of the liver-specific extracellular matrix (LS-ECM) and become more resistant to the treatment with molecular Doxorubicin (Dox) and Dox-loaded mesoporous silica nanoparticles than in monolayer cultures. When grafted on CE chorioallantoic membrane, LS-ECM-based TECs induced angiogenic switch. These findings may have important implications for the diag-nosis and treatment of TNBC. The methodology established here is scalable and adaptable for pharmacological testing and cancer biology research of various metastatic and primary tumors.

Mountain and pastoralist societies around the world have for centuries sustained their livelihoods and cultures by accumulating specialist knowledge about their local and regional socio-ecological environments. Developing traditional knowledge and customary practices takes time, sometimes spanning across generations. As macro-level changes to social and natural environment are now taking place, such as globalization and climate change, local communities could potentially also benefit from complementary, suitably adapted educational opportunities for sustainable development. However, access to education has often required moving to urban centres, which can weaken community structures and cohesion, and could also foster increased dependence on external specialists, providers or decision-makers. Careful introduction of emerging Educational Technologies could alleviate and possibly reverse such trends as mobile Internet access spreads to remote areas. This paper examines the role of education in sustainable development and specifically explores the potential for two educational innovations, open badges and blockchain, to provide a new construct for transformation in sustainable development amongst mountain and pastoralist societies. These technologies could not only facilitate education through online distance learning, but also allow geographically remote populations to highlight the value of their traditional knowledge and to engage more comprehensively in their changing worlds.

Pseudopterosin, produced by the sea whip of the genus Antillogorgia, possesses a variety of promising biological activities including potent anti-inflammatory effects. However, few studies examined pseudopterosin in the treatment of cancer cells and, to our knowledge, the ability to inhibit triple negative breast cancer (TNBC) proliferation or invasion has not been explored. Thus, we evaluated the as yet unknown mechanism of action of pseudopterosin: Pseudopterosin was able to inhibit proliferation of TNBC. Interestingly, analyzing breast cancer cell proliferation after knocking down glucocorticoid receptor α (GRα) revealed that anti-proliferative effects of pseudopterosin were significantly inhibited when GRα expression was reduced. Furthermore, pseudopterosin inhibited invasion of MDA-MB-231 3D tumor spheroids embedded in an extracellular-like matrix. Remarkably, the knockdown of GRα in 3D tumor spheroids revealed increased ability of cells to invade the surrounding matrix. In a co-culture, encompassing peripheral blood mononuclear cells (PBMC) and MDA-MB-231 cells, production of interleukin 6 (IL-6) and interleukin 8 (IL-8) significantly increased compared to monoculture. Notably, pseudopterosin proved to block cytokine elevation, representing key players in tumor progression, in the co-culture. Thus, our results reveal pseudopterosin treatment as a potential novel approach in TNBC therapy.

Oxidative stress (OS) is involved in the pathogenesis of retinal neurodegenerative diseases like age-related macular degeneration (AMD) and diabetic retinopathy (DR) and an important target of therapeutic treatments. New therapeutics are tested in vivo despite limits in transferability and ethical concerns. Retina cultures using human tissue can deliver critical information and significantly reduce the number of animal experiments along with increased transferability. We cultured up to 32 retina samples derived from one eye, analyzed models’ quality, induced OS, and tested efficiency of antioxidative therapeutics. Bovine, porcine, rat, and human retinae were cultured in different experimental settings for 3-14 d. OS was induced by high-glucose or hydrogen peroxide (H2O2) and treated by Scutellarin, pigment epithelium-derived factor (PEDF), and/or granulocyte macrophage-colony stimulating factor (GM-CSF). Tissue morphology, cell viability, inflammation, and glutathione level were determined. Retina samples showed only moderate necrosis (23.83±5.05 increased to 27.00±1.66 AU PI-staining over 14 d) after 14 days in culture. OS was successfully induced (reduced ATP content of 288.3±59.9 vs. 435.7±166.8 nM ATP in controls); antioxidants reduced OS-induced apoptosis (from 124.20±51.09 to 60.80±319.66 cells/image after scutellarin-treatment). Enhanced mammalian animal and human retina cultures allow reliable, highly transferable research on OS-triggered age-related diseases and pre-clinical testing during drug development.

Gene electrotransfer is one of the main non-viral methods for intracellular delivery of plasmid DNA, wherein pulsed electric fields are used to transiently permeabilize the cell membrane allowing enhanced transmembrane transport. By localizing the electric field over small portions of the cell membrane using nanostructured substrates, it is possible to increase considerably the gene electrotransfer efficiency while preserving cell viability. In this study, we design an electrotransfer approach based on commercially available cell culture inserts with polyethylene-terephthalate (PET) porous substrate. We first use multiscale numerical modelling to determine the pulse parameters, substrate pore size, and other factors that are expected to result in successful gene electrotransfer. Based on numerical results we design a simple device combining an insert with substrates containing pores with 0.4 um and 1.0 um diameter, a multiwell plate, and a pair of wire electrodes. We test the device in three mammalian cell lines and obtain transfection efficiencies similar to those achieved with bulk electroporation, but with low-voltage pulses that do not require the use of expensive electroporators. Our combined theoretical and experimental analysis calls for further systematic studies that will investigate the influence of substrate pore size and porosity on gene electrotransfer efficiency and cell viability.

In this article, you will get acquainted with ancient settlements in the Bukhara region and settlements found at the foot of the Zarafshan River. In addition, the settlements of urbanized peasant and artisan families belonging to the ancient Sugdian culture are also covered. Just as Egypt was called "the gift of the Nile" in ancient times, Buhoro oasis can also be called the gift of the Zarafshan river. Because this land on the banks of the river itself, first of all, as a result of thousands of years of Zarafshan was created due to the turbidity that flowed. The innumerable streams, lakes and ponds formed by its floods played a major role in the formation of the nature of the oasis and its development. It is known that when the ice age ended and the weather started to warm, the thick glaciers and eternal snows that covered the mountain ranges of Turkestan, Khisor and Zarafshan melted, and huge floods formed from them, roared down the Zarafshan river. His depicted floods of those times reminded of the legendary "Noah's flood". At that time, Zarafshan flowed over a very wide area in its valley. Its right bank washes the Payarik and Khatirchi hills in the present Samarkand region, and the left bank washes the Pastdargom and Kattakurgan hills. Not a single drop of this large stream was wasted in the upper and middle parts of the Zarafshan valley and was used for farming and agriculture, it broke through the narrow Khazora dar band near the present city of Navoi and flowed into the cone-shaped Bukhoro oasis. Ancient Khitfar (Vobkent-Darya), Rudnzor (Shakhrud) in Bukhara oasis. Korakuldaryo, Mokhondaryo and Taikir formed several branches and flowed along the entire voka. At that time, the valley part of Bukhoro region was formed from the lakes and swamps formed by the flood waters of Zarafshan.They are covered with thick bushes, thickets and orchards. A wealth of information has been collected on the history of the ancient nature of the people of the country and the development of its unique culture, as well as the formation of self-interested cattle breeding and farming farms, as well as the emergence of large and small villages and cities. The data serve as the main source not only for the history of the emergence of the city of Bukhara, but also for the study of the emergence of the process of urbanization in the Zara Valley and its specific characteristics.

Tumor growth, progression, and therapy resistance are crucial factors in the prognosis of cancer. Properties of three-dimensional tumor-like organoids (tumoroids) more closely resemble in vivo tumors compared to two-dimensionally cultured cells and are therefore effectively used for assays and drug screening. We here established a repurposed drug for novel anticancer research and therapeutics using a tumoroid-based screening system. We screened 6 pharmacologically active compounds by using an original tumoroid-based multiplex phenotypic screening system with matrix metalloproteinase 9 (MMP9) promoter-driven fluorescence reporter for the evaluation of both tumoroid formation and progression. The effects of one of the hit compounds were examined on tumor formation and progression in vitro and in vivo. Antiparkinson drug benztropine was the most effective compound uncovered by the screen. Benztropine significantly inhibited in vitro tumoroid formation, cancer cell survival, and MMP9 promoter activity. Benztropine also reduced the activity of oncogenic signaling transducers and trans-activators for MMP9, including STAT3, NF-κB, and β-catenin, and properties of cancer stem cells / cancer-initiating cells. Benztropine and GBR-12935 directly targeted the dopamine transporter DAT/SLC6A3, whose genetic alterations such as amplification were correlated with poor prognosis for cancer patients. Benztropine also inhibited tumor growth, circulating tumor cell (CTC) number, and rate of metastasis in a tumor allograft model in mice. In conclusion, we propose the repurposing of benztropine for anticancer research and therapeutics that can suppress tumor progression, CTC, and metastasis of aggressive cancers by reducing key pro-tumorigenic factors.

Glial cells provide physical and chemical support and protection for neurons. Secreted neurotrophic factors, scaffolds and vesicles regulate in addition to normal homeostasis, repair following neural tissue injury or degeneration. We previously demonstrated that human CNS glia have revascularization capacity by inducing endothelial cell sprouting and microchanneling driven by glial cell secreted components. The clinical potential of using secreted factors and vesicles generated by glial cells in conditioned media was shown by their re-normalization of pathological blood vessels and 3D tissue wound healing and remodeling capacity. To identify secreted components of glial cells and how CNS and PNS glial cells differentially regulate neural tissue homeostasis, disease and injury response, we developed a novel culture method for various types of glial cells that allow for isolating and characterizing secreted glial components, free of animal factors and contaminants normally associated with serum-based culture conditions. Our culture method will enable to identify secreted components using ultrasensitive RNA, glycoconjugate and protein analysis technologies. The characterization of glial secreted factors involved microchanneling may help in development of new clinical therapies for glial and neural cells or axons to migrate and reform connections.

The prevalence of internet gaming disorders (IGD) is considerably high among youth, especially with social isolation imposed by the ongoing COVID-19 pandemic. IGD adversely affects mental health, quality of life, and academic performance. The Internet Gaming Disorder Scale (IGDS9-SF) is designed to detect IGD according to DSM-IV diagnostic criteria. However, inconsistent results are reported on its capacity to diagnose IGD evenly across different cultures. To ensure the suitability of the IGDS9-SF as a global measure of IGD, this study examined the psychometric properties of the IGDS9-SF in a sample of Sri Lankan university students (N = 322, mean age =17.2 ± 0.6, range = 16-18 years, 56.5% males) and evaluated its measurement invariance across samples from Sri Lanka, Turkey, Australia, and the USA. Among Sri Lankan students, a unidimensional structure expressed good fit, invariance across different groups (gender, ethnicity, income, etc.), adequate criterion validity (strong correlation with motives of internet gaming, daily gaming duration, and sleep quality), and good reliability (alpha = 0.81). Males and online multiplayers expressed higher IGD levels, greater time spent gaming, and more endorsement of gaming motives (e.g., Social and Coping) than females and offline players. Across countries, the IGDS9-SF was invariant at the configural, metric, and scalar levels, albeit strict invariance was not maintained. The lowest and highest IGD levels were reported among Turkish and American respondents, respectively. In conclusion, the IGDS9-SF can be reliably used to measure IGD among Sri Lankan youth. Because the scale holds scalar invariance across countries, its scores can be used to compare IGD levels in the studied countries.

In this paper I tentatively answer 50 questions sampled from a pool of over 10,000 weekly questions formulated by students in a course entitled “The Self”. The questions pertain to various key topics about self-processes, such as self-awareness, self-knowledge, self-regulation, self-talk, self-esteem, and self-regulation. The students’ weekly questions and their answers highlight what is currently know about the self. Answers to the student questions also allow for the identification of some recurrent lessons about the self. Some of these lessons include: all self-processes are interconnected (e.g., prospection depends on autobiography), self-terms must be properly defined (e.g., self-rumination and worry are not the same), inner speech plays an important role in self-processes, controversies are numerous (are animals self-aware?), measurement issues abound (e.g., self-reflection as an operationalization of self-awareness), deficits in some self-processes can have devastating effects (e.g., self-regulatory deficits may lead to financial problems), and there are lots of unknowns about the self (e.g., gender differences in Theory-of-Mind).

The literature lacks consensus on the minimum microbial density required for diagnosing urinary tract infections (UTIs). This study categorized the microbial densities of urine specimens from symptomatic presumptive UTI patients and correlated them with detected levels of immune response biomarkers NGAL, IL-8, and IL-1β. The objective was to identify changes in immune response at different microbial densities and determine an optimal threshold for diagnosing symptomatic UTIs. Midstream clean-catch urine samples were analyzed for microbial identification and quantification using SUC and M-PCR, while NGAL, IL-8, and IL-1β levels were measured with ELISA. NGAL, IL-8, and IL-1β levels were all significantly higher in symptomatic subjects with microbial densities ≥ 10,000 cells/mL by M-PCR (p < 0.0069) or equivalent CFUs/mL by SUC (p < 0.0104) compared to samples with no detectable microbes. With both PCR and SUC, a consensus of two or more elevated biomarkers correlated well with microbial densities > 10,000 cells/mL or CFU/mL, respectively. The association between ≥ 10,000 cells and CFU per mL with elevated biomarkers in symptomatic patients suggests that this lower threshold may be more suitable for diagnosing UTIs. These findings challenge the widely used threshold of 100,000 CFU per mL, which may overlook many active UTI cases.

The Environmental interactions contributed to the processes of settlement and development of the first settlers at southern Peru 15,000 BC. The determination of this process is based on paleoclimatic studies at southern Peru and ethnoarchaeological evidence. The determination of this process is based on paleoclimatic studies of southern Peru and ethnoarchaeological evidence; establishing as a result of this analysis the hypothesis of environmental interaction and territorial occupation of the first hunter-gatherer populations until their consolidation, dominating the geographical space from Tacna. The Disruptive disaster events and their effects on climate during the Peruvian Paleolithic resulted in the origin of local societies that shaped the Tiawanaku societies of the early horizon at Southern Peru and Northern Chile.

Antimicrobial peptides provide a powerful toolkit for combating multidrug resistance. Combatting eukaryotic pathogens is complicated by the fact, the intracellular drug targets in the eukaryotic pathogen are frequently homologs of cellular structures of vital importance in the host organism. The entomopathogenic bacteria, (symbionts of entomopathogenic nematode species), release a series of non-ribosomal templated antimicrobial peptides, some may be potential drug candidates. The ability of an entomopathogenic nematode /entomopathogenic bacterium symbiotic complex to survive in a given polyxenic milieu is a coevolutionary product, explaining that those gene complexes which are responsible for the biosynthesis of different non-ribosomal templated antimicrobial protective peptides (including those which are potently capable to inactivate the protist mammalian -pathogen Leishmania and the gallinaceous bird pathogen Histomonas) are co-regulated. Our approach is based on comparative antimicrobial bioassays of the culture media of the wild-type and regulatory mutant strains. We concluded that Xenorhabdus budapestensis and X. szentirmaii are excellent sources of non-ribosomal templated antimicrobial peptides, that are efficient antagonists of the mentioned pathogens. Data on selective cytotoxicity of different cell-free culture media encourages us to forecast that the recently discovered “easy-PACId” research strategy is suitable for constructing EPB strains producing and releasing single, harmless, non-ribosomal templated antimicrobial peptides with considerable drug, (probiotic) -candidate potential.

The goal of this study was to determine the relative susceptibility of four common equine wound pathogens to ten different types of antimicrobial agents based on minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Additionally, to describe the comparison of bactericidal activity of medical grade Manuka honey, local honey, and commercial, food-grade honey to other commonly used wound dressings. The objective is to provide veterinary practitioners with comparative data on the use of a variety of antimicrobial dressings for inhibiting growth of common wound bacteria. MIC and MBC for Manuka, store, and local honeys were comparable to those of sterile gauze, sugar, and hypertonic saline. Across bacterial species, local honey proved to have more bactericidal activity when compared to Manuka and commercial, food-grade honey. The MIC and MBC for PHMB gauze and foam was consistently at a higher dilution compared to the other antimicrobials. The majority of antimicrobials exhibited stronger inhibitory and bactericidal activity against a Streptococcus zooepidemicus isolate obtained from a wound compared to other bacteria that were ATCC lab acclimated. Additional research for in-vivo applications needs to be done to see whether differences exist in effective wound management.