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 study is based on the notion that big data predictive analytics is important for developing strategic alliances performance of companies. this study investigates the relationship between big data predictive analytics, big data culture, and competitive strategies 'techniques were adopted, such as descriptive statistics, correlation,regression, etc. using SPSS and SmartPLS statistical software. Hypotheses were tested with bootstrapped analysis using SEM (through SmartPLS). The study developed a structural equation model by using the SEM analysis. The results of the SEM analysis suggested the hypothesized model of the study was valid. The results supported all the hypotheses of the study.Through empirical analysis, demonstrated the conclusion is that the big data predictive analytics has a positive and significant relationship with strategic alliance performance

The objective of this study was to analyze the in vitro stability and toxicity of liposomes containing guarana in skin cell lines. The liposomes were produced by the reverse phase evaporation method containing 1 mg/mL guarana. The stability of the liposomes was evaluated by physical-chemical parameters for up to 90 d using three different storage conditions. The cytotoxicity of guarana (GL), liposomes (B-Lip), and guarana-loaded liposomes (G-Lip) was evaluated on spontaneously immortalized human keratinocyte cell lines (HaCaT), murine swiss albino fibroblasts (3T3), and human fibroblast (1BR.3.G). The evaluation was performed using cellular viability analysis. The techniques used were 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red capturing (NRU) and the analyses were conducted after 24, 48, and 72 h exposure of these cells to the different treatments. The G-Lip exhibited physical-chemical stability for 60 d when the samples were stored in a refrigerator. The GL, B-Lip, and G-Lip demonstrated low cytotoxicity in the three cell cultures tested since a low reduction in cell viability was only observed at the highest concentrations. In addition, greater cell damage was observed for B-Lip; however, guarana protects cells from this damage. Thus, G-Lip structures can be considered as promising systems for topical applications.

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.

The brown seaweed Alaria esculenta is the second most cultivated species in Europe and it is therefore of interest to expand its application in developing food products. In this study, a lactic acid bacteria consortium (LAB consortium) consisting of three Lactiplantabacillus plantarum strains (relative abundance ~ 94%) and a minor amount of a Levilactobacillus brevis strain (relative abundance of ~ 6%) was investigated for its ability to ferment carbohydrates available in brown seaweed. The consortium demonstrated ability in fermenting glucose, mannitol, galactose, mannose, and xylose, of which glucose and mannitol were the most favored substrates; no growth was observed on fucose, mannuronic and guluronic acid. The consortium used different pathways for carbohydrate utilization and produced lactic acid as the main metabolite. In glucose fermentations, only lactic acid was produced, but use of mannitol as carbohydrate source, resulted in co-production of lactic acid, ethanol and succinate. Xylose fermentation resulted in acetate production. The consortium was also able to utilize laminari-oligosaccharides (DP 2-4), obtained after enzymatic hydrolysis of laminarin, and produced lactic acid as metabolite. The consortium could grow directly on A. esculenta, resulting in a pH decrease to 3.8 after 7 days of fermentation. Incubation of the same seaweed at corresponding conditions without inoculation resulted in spoilage of the seaweed by endogenous bacteria.

This qualitative research study explores the experiences of three Emirati women who have achieved significant success in their higher education institutes in the (UAE). The study aims to understand how childhood and adolescent experiences shape the leadership development of these women, both positively and negatively. The study also includes interviews with family members and close colleagues of the participants to gain additional perspectives. The findings the importance of family support, role models, and mentors in shaping the leadership development of women in the UAE. The study also sheds light on the challenges and barriers that women face in leadership positions, including gender bias and discrimination. Overall, the study contributes to a deeper understanding of the experiences of Emirati women in leadership positions and provides insights into how to support their leadership development.

Respiratory tract epithelium infection plays a primary role in Nipah virus (NiV) pathogenesis and transmission. Knowledge about infection dynamics and host responses to NiV infection in respir-atory tract epithelia is scarce. Studies in non-differentiated primary respiratory tract cells or cell lines indicate insufficient interferon (IFN) responses. However, studies are lacking to determin-ing complex host response patterns in differentiated respiratory tract epithelia to understand NiV replication and spread in swine. Here we characterized infection and spread of NiV in differenti-ated primary porcine bronchial epithelial cells (PBEC) cultivated at the air-liquid-interface (ALI). After the initial infection of only a few apical cells, lateral spread for 12 days with epithelium disruption was observed without releasing substantial amounts of infectious virus from the api-cal or basal sides. Deep time course proteomics revealed pronounced upregulation of genes re-lated to type I/II- IFN, immunoproteasomal subunits, TAP-mediated peptide transport and MHC I antigen presentation. Spliceosomal factors were downregulated. We propose a model in which NiV replication in PBEC is slowed by a potent and broad type I/II-IFN host response with con-version from 26S proteasomes to immunoproteasomal antigen processing and improved MHC I presentation for adaptive immunity priming. NiV induced cytopathic effects could reflect the focal release of cell-associated NiV, which may contribute to efficient airborne viral spread between pigs.

Digital microfluidics (DMF) holds great potential for the alleviation of laboratory procedures in assisted reproductive technologies (ARTs). Electrowetting on dielectric (EWOD) provides dynamic culture conditions in vitro that may better mimic the natural embryo microenvironment. Thus far, EWOD microdevices have been proposed for in vitro gamete and embryo handling in mice and for analyzing the human embryo secretome. This article presents the development of the first microfluidic chip utilizing EWOD technology designed for the manipulation of bovine embryos in vitro. The prototype sustains the cell cycles of embryos manipulated individually on the chips during in vitro culture (IVC). Challenges related to the chip fabrication as well as to its application during bovine embryo IVC in accordance with the adapted on-chip protocol are thoroughly discussed, and future directions for DMF in ARTs are indicated.

Viruses with encephalitogenic potential can cause neurological conditions of clinical and epidemiological importance, such as Saint Louis encephalitis virus, Venezuelan equine encephalitis virus, Eastern equine encephalitis virus, Western equine encephalitis virus, dengue virus, Zika virus, Chikungunya virus, Mayaro virus and West Nile virus. The objective of the present study was to determine the number of arboviruses with neuroinvasive potential isolated in Brazil that corresponds to the collection of viral samples belonging to the Section of Arbovirology and Hemorrhagic Fevers (SAARB/IEC) of the Laboratory Network of National Reference for Arbovirus Diagnosis from 1954 to 2022. A total of 1,090 arbovirus samples with encephalitogenic potential were isolated from mice. From a total of 7,054 samples intended for cell culture. The isolates from mosquitoes totaled 659 viruses. The emergence of new arboviruses may be responsible for diseases still unknown to humans, making the Amazon region a hotspot for infectious diseases due to its fauna and flora species characteristics. The detection of circulating arboviruses with the potential to cause neuroinvasive diseases is constant, which justifies the continuation of active epidemiological surveillance work that offers adequate support to the public health system regarding the virological diagnosis of circulating arboviruses in Brazil.

Natural astaxanthin is a precious substance obtained from some organisms such as microalgae. This plant has many benefits for humans, so research into its cost-effective and economical production has recently increased. For this purpose, some methods such as the use of different culture media, gene engineering, different stresses, nanoparticles, bio acids, and phytohormones are important. Accordingly, this review study was conducted to demonstrate the effect of the factors mentioned above for the high production of astaxanthin in microalgae, especially Haematococcus pluvialis (H.p).

This prospective single-center study was designed to assess the clinical utility of the FilmArray® blood culture identification (BCID) panel for improving the diagnostic accuracy in neonatal sepsis. Results obtained using the FilmArray® BCID panel were correlated with results of blood culture in all consecutive neonates with suspicion of early-onset (EOS) and late-onset sepsis (LOS) attended in our service over a 2-year period. A total of 102 blood cultures from 92 neonates were included, 69 (67.5%) in cases of EOS and 33 (32.3%) in LOS. The FilmArray® BCID panel was performed in negative culture bottles at a median of 10 hours of blood culture incubation (range 7.5-20 hours), without differences by the type of sepsis. The FilmArray® BCID panel showed a 66.7% sensitivity, 100% specificity, 100% positive predictive value, and 95.7% negative predictive value. There were four false-negative cases, three of which of Streptococcus epidermidis in neonates with LOS and one case of Granulicatella adiacens in one neonate with EOS. We conclude that the use of the FilmArray® BCID panel in negative blood cultures from neonates with clinical suspicion of sepsis is useful in decision-making of starting or early withdrawal of empirical antimicrobials because of the high specificity and negative predictive values of this assay.

Digital transformation of companies is often referred to as a future track and a strategic path towards ensuring the survival and sustainability of companies. In this paper, we aim to outline this transformation process in Tunisian companies and identify its driving factors and finally explain its key success factors. To this end, we examine a sample of 70 companies operating in all economic sectors. The results indicate that such a digital strategy has a significant and a positive effect on the success of digital transformation of companies. Leadership has an effect at a low level of digital maturity. On the other hand, corporate culture does not have a significant effect on digital transformation. These results contribute to explaining this emerging phenomenon by focusing on the driving factors as well as the factors that contribute to its success.

Concentration and preservation of lactic acid bacteria (LAB) starter cultures for food production guarantee the long-term delivery of stable cultures in terms of viability and functional activity. One method that has commonly been used to prepare dried starter cultures for food applications is freeze-drying. During freeze-drying, the bacterial cells are exposed to stresses such as freezing, drying, and long-term exposure to low-water activity. Additionally, another added stress is introduced during the rehydration process. The purpose of this study was to evaluate the survival rates and fermentation performance of three (3) freeze-dried lactic acid bacterial cultures previously isolated from Ghanaian traditional fermented milk. LAB cultures, i.e., Lactobacillus delbrueckii, Lactococcus lactis and Leuconostoc mesenteroides were frozen in the chamber of a Telstar (Lyoquest) laboratory freeze dryer for 10 hrs at -55 °C (as single and combined cultures) using skimmed milk and cassava flour as cryoprotectants held in plastic or glass cryovial. For viability during storage, freeze-dried LAB cultures were stored in a refrigerator (4 °C) and at room temperature (25 °C) for 4-weeks. The survival of freeze-dried cultures was determined by growth kinetics at 600nm (OD₆₀₀). The performance of freeze-dried LAB cultures after 4-weeks storage was determined by their growth and acidification of milk during yogurt fermentation and consumer sensory evaluation of fermented milk using a 9-point hedonic scale. The survival rates for LAB ranged between 60.11% and 70.91% following freeze-drying. For single cultures, the highest survival was recorded for Lactobacillus delbrueckii (L12), whereas for combined cultures, the highest survival was observed for Lactococcus lactis (L3) combined with Lactobacillus delbrueckii (L12). During the fermentation process all the freeze-dried LAB cultures were able to acidify yogurt to a pH below 4, while yogurt produce from the spontaneous fermentation was characterized by low acidification. Yogurt fermented with freeze-dried lactic acid bacteria cultures, either single or combined strains, showed improved acceptability as compared to the spontaneously fermented yogurt. The consumer acceptability results showed that yogurts produced with combined starter culture of Lactoccus lactis and Lactobacillus delbrueckii or single culture of Lactococcus lactis were the most preferred products with Lactococcus lactis and Lactobacillus delbrueckii possessing high survival rates and high consumer acceptability in yogurt production. These findings are crucial and can be adopted for large-scale production and commercialization of yogurt production, however, in-depth investigation on the effects of freeze-drying and long-term storage on survival and performance of selected LAB cultures are needed.

The genus Musa spp. contains commercially important fleshy fruit producing plants including plantains and bananas with a strong potential of providing food security and source of revenue to farmers. Occasionally, plantlets are often not available to fulfill the supply demand of farmers particularly in Caribbean region. Concerns with the quality of vegetative tissues along with the possibility of the transmission of phytopathogens makes availability of plantlets limited to farm-ers. Micropropagation of plantains offers an alternative to producing large number of in-vitro plantlets. However, conventional methods of micropropagation techniques require high produc-tion costs and are labor-intensive. Recently, Temporary Immersion Bioreactor (TIB) has emerged as an alternative to conventional micropropagation methods. Our work utilized SEM (Scanning Electron Microscope), molecular and biochemical tools (qRT-PCR and ICP-OES) to characterize and compare the morphological, elemental composition, and photosynthetic gene expression of plantains cultured on TIB. Additionally, morphological features of growth and propagation rates were analyzed for comparing outputs obtained from temporary immersion bioreactor with conventional micropropagation (CM) techniques. Results showed higher growth and multiplication rates for plantlets cultivated in TIB. Gene expression analysis of selected photosynthetic genes demonstrated high transcript abundance of phosphoenol pyruvate carboxylase (PEPC) in plantain tissues obtained by TIB. Elemental composition analysis showed higher content of iron in plantains grown in TIB suggesting a potential correlation with PEPC expression. These results demonstrate the potential of TIB to be an efficient method to produce healthy in-vitro plantains.

This paper looks at key elements of global culture that are driving a new paradigm shift in museums causing them to question their raison d'être, their design and physical space, recognizing the need to accommodate visitor interaction and participation, and to reprioritize institutional outcomes and goals reexamining their priorities. As heritage sharing in online spaces reaches across national, political, and social boundaries on platforms and networks, this has been driven by museum engagement with Internet life during the pandemic. Museum relationships and interactions with communities both local and global, continue to challenge core values and precepts, leading to radical changes in how museums define their roles and responsibilities. In this new cultural landscape, museums are responding to human digital identity in a tidal wave of human interactions on the Internet, from social media to online sharing of images and videos. This is revealing shared perspectives on cultural conflict as being tied to freedom of expression of one’s heritage embedded in digital identity.

Several studies have highlighted the importance and evolution of process safety leadership and culture in various industries. However, none has focused on the palm oil milling industry yet. This paper critically reviews the latest developments in the palm oil milling process and unit operations leading to process safety concerns. It also discusses the Principle of 3C that is applied to explain repeat accidents and the four-level safety culture in the palm oil milling industry. For this purpose, the author presents case studies of two key palm oil companies in Malaysia. Overall, this paper offers guidelines to leaders in the palm oil milling industry about the required process safety leadership and culture to be understood in order to improve their safety outcomes.

Ovine footrot is a complex multifactorial infectious disease, causing lameness in sheep with major welfare and economic consequences. Dichelobacter nodosus is the main causative bacterium, however, footrot is a polymicrobial disease with Fusobacterium necrophorum, Mycoplasma fermentans and Porphyromonas asaccharolytica also associated. There is limited understanding of the host response involved. Proinflammatory mediators interleukin (IL)-1β and C-X-C Motif Chemokine Ligand 8 (CXCL8) have been shown to play a role in the early response to D. nodosus in dermal fibroblasts and interdigital skin explant models. To further understand the response of ovine skin to bacterial stimulation, and to build the understanding of the role of the cytokines and chemokines identified in transcriptomic data, primary ovine interdigital fibroblasts and keratinocytes were isolated, cultured and stimulated with lipopolysaccharide (LPS), D. nodosus or F. necrophorum in the presence and absence of M. fermentans, whilst measuring mRNA expression and protein release of CXCL8 and, IL-1β. Stimulation with LPS, D. nodosus or F. necrophorum resulted in increased transcript levels of IL-1β and CXCL8 in M. fermentans free cells, however, only an increase in CXCL8 protein release was observed. No IL-1β protein release was detected despite increases in IL-1β mRNA, suggesting the signal for intracellular pre-IL-1β processing may be lacking when culturing primary cells in isolation. Keratinocytes and fibroblasts naturally infected with M. fermentans showed little response to LPS, a range of D. nodosus preparations or heat-inactivated F. necrophorum. Primary single cell culture models complement ex vivo organ culture models to study different aspects of the host response to D. nodosus. Ovine keratinocytes and fibroblasts infected with M. fermentans had a reduced response to experimental bacterial stimulation. However, in the case of footrot where Mycoplasma spp. are associated with diseased feet, this natural infection gives important insights into the impact of multiple pathogens on the host response.

Standard cell culture is routinely performed at supraphysiological oxygen concentrations (~18% O2). Conversely, oxygen levels in most tissues range from 1%–6% (physioxia). Such hyperoxic conditions can alter reactive oxygen species (ROS) production, energy metabolism, mitochondrial network dynamics, and response to drugs and hormones. The aim of this project was to investigate the transcriptional response to different oxygen levels and whether it is similar across cell lines, or cell-line specific. Using RNA-seq, we performed differential gene expression and functional enrichment analyses in four human cancer cell lines, LNCaP, Huh-7, PC-3, and SH-SY5Y cultured at either 5% or 18% oxygen for 14 days. We found that oxygen levels affected transcript abundance of hundreds of genes, with the affected genes having little overlap between cell lines. Functional enrichment analysis also revealed different processes and pathways being affected in each cell line. Interestingly, we found that the top differentially expressed genes are involved in cancer biology. Further, we observed several hypoxia-inducible factor (HIF) targets upregulated at 5% oxygen, suggesting a role of HIF at physiological oxygen conditions. Finally, oxygen strongly induced transcription of mitochondrial genes in most cell lines, in a cell-type specific manner too. We conclude that cellular response to oxygen is widely cell-type specific, emphasizing the importance of maintaining physioxia in cell culture.

The efficiency of rice cum fish culture study was conducted in different areas of Tangail district with those farmers who cultivate rice and fish together at a time. The study was conducted along 60 farmers of different areas in Ghatail upazilla. The primary data were collected through face-to-face interviews and secondary data were collected through different reputed journals, newspapers, authentic portals and so on. Data collection was carried out during the period from November 2021 to May 2022. Both tabular and functional analyses were performed to achieve the specific objectives of the study. NPV, BCR and IRR method was used to estimate the profitability and a multiple regression model was used to find out the factors motivating farmers' decision on rice cum fish cultivation. SWOT analysis was done to find out the internal and external-positive and negative factors faced by the farmers in rice cum fish cultivation. About 40% of farmers are under the age of 30 and most of them got at least primary education. Agriculture was the main occupation of the respondents and fish cultivation was the secondary. The highest annual income among the sample growers came from the production of rice cum fish culture. The BCR of the production is upbeat and 2.09 which indicates that the cultivation is profitable.

Cancer invasion through basement membranes represents the initial step of tumor dissemination and metastasis. However, little is known about how human cancer cells breach basement membranes. Here, we used a 3-dimensional in vitro invasion model consisting of cancer spheroids encapsulated by a basement membrane and embedded in 3D collagen gels to visualize the early events of cancer invasion by confocal microscopy and live-cell imaging. Human breast cancer cells generated large numbers of basement membrane perforations, or holes, of varying sizes that expanded over time during cell invasion. We used a wide variety of small molecule inhibitors to probe the mechanisms of basement membrane perforation and hole expansion. Protease inhibitor treatment (BB94), led to a 63% decrease in perforation size. After myosin II inhibition (blebbistatin), basement membrane perforation area decreased by only 15%. These treatments produced correspondingly decreased cellular breaching events. Interestingly, inhibition of actin polymerization dramatically decreased basement membrane perforation by 80% and blocked invasion. Our findings suggest that human cancer cells can primarily use proteolysis and actin polymerization to perforate the BM and to expand perforations for basement membrane breaching, with a relatively small contribution from myosin II contractility.

Leishmania parasites present astonishing adaptative abilities that represent a matter of life or death within disparate environments during the heteroxenous parasite life cycle. From an evolutionary perspective, organisms develop methods of overcoming such challenges. Strategies that extend beyond the genetic diversity have been discussed and include variability between parasite cells during the infections of their hosts. The occurrence of Leishmania subpopulation fluctuations with variable structural genomic contents demonstrates that a single strain might shelter the variability required to overcome inconsistent environments. Such intrastrain variability provides parasites with an extraordinary ability to adapt and thus survive and propagate. However, different perspectives on this evolution have been proposed. Strains or species living in the same environment can cooperate but also compete. These interactions might increase the replication rate of some parasites but cause the loss of more aggressive competitors for others. Adaptive responses to intra- and interspecific competition can evolve as a fixed strategy (replication is adapted to the average genetic complexity of infections) or an optional strategy (replication varies according to the genetic complexity of the current infection). This review highlights the complexity of interspecies and intrastrain interactions among Leishmania parasites as well as the different factors that influence this interplay.

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.

Human pluripotent stem cells (hPSCs) have become a powerful tool to generate various kinds of cell types comprising the human body. Recently, organoid technology emerged as a platform to build a physiologically relevant tissue-like structure from the PSCs, which provides a more relevant three-dimensional microenvironment to the actual human body than the conventional monolayer culture system for transplantation, disease modeling, and drug development. Although it holds so many advantages, the organoid culture system still has various problems related to culture methods, which became a challenge to get similar physiological properties to their original tissue counterparts. Here, we discuss the current development of organoid culture methods, including the problem that may arise from the currently available culture systems as well as the possible approach to overcoming the current limitation and improving their optimum utilization for translational application purposes.

Port cities’ sustainable development can start from innovation in maritime culture to build new urban visions based on the goals of Agenda 2030 and oriented towards local and international cooperation. In the international debate innovative strategies on cultural heritage enhancement contaminate the research and production contexts of ports. In addition, numerous cities have implemented creative and cultural responses to climate change and environmental sustainability. Creativity and cultural heritage enhancement can guide the definition of new trajectories of sustainable urban development, particularly in port-city interaction areas. In Europe, port-city interaction areas have been transformed into laboratories of cultural and creative experimentation for the sustainable management of cultural heritage and the urban quality of public spaces. In this perspective, the paper, starting from the studies developed on the main measurement frameworks of creative cities and sustainable development policies, aims to investigate the possibility of developing a "Port-cities Creative Heritage Enhancement" approach to assess and plan possible cultural and creative transformations of historical-architectural buildings, industrial archaeologies and symbolic urban spaces in the port-city interaction areas of Naples.

Objectives: The promising specific growth regulators are employed tissue culture of the bamboo species. The specific natural hardening mixture supports the acclimatization and adaptation of the protected cultivation. The bamboo species contribute to the carbon sequestration and stabilization of the environment. Findings: The growth regulator 2, 4-D encourages callus induction and growth regulators NAA + TDZ, BAP + Kinetin + Gelrite, BAP + Kinetin + Coconut water + myo-inositol, NAA and TDZ impose plant regeneration in the Bambusa species. The growth regulator of 2,4-D, 2,4-D + Kinetin, 2,4-D + BA, 2,4-D + BAP and 2,4-D + NAA + BAP produces callus induction in the Dendrocalamus species. The growth regulator of TDZ, BA and IBA + Coumarin + Putrescine, BAP + GA3, NAA + Kinetin, BA + IBA, BA + Kinetin + NAA, NAA + Kinetin, Kinetin + IAA has facilitated shoot regeneration in the Dendrocalamus species with the specific period. The growth regulator of BA and TDZ; BAP + TDZ, IBA + Coumarin + Putrescine, BAP + GA3, NAA + Kinetin, BA + IBA, BA + Kinetin + NAA, NAA + Kinetin, Kinetin + IAA develops root regeneration with unambiguous time in the Dendrocalamus species. The growth regulator of BAP and Kinetin + BAP imposes shoot regeneration and IBA and IBA + Coumarin + sucrose conducts root regeneration in the edible bamboo. The natural hardening materials of cocopeat, vermicompost, perlite, cow dung, FYM, compost, soil & garden soil and humus soil, perlite, and FYM recommend in the acclimatization and adaptation of the Bambusa species and Dendrocalamus species respectively. Novelty: The standard growth regulators and hardening mixtures impose tissue culture, acclimatization and adaptation in the bamboo species. The bamboo species involves in managing carbon sequestration, biogeochemical cycle and the environment.

Perennial cropping play vital roles in regulating soil carbon sequestration and thus mitigating climate change. However, how perennial cropping affects soil microbial community remains elusive. Using a field investigation, this study was conducted to examine the effects of mugwort cropping along a chronosequence (that is, wheat-maize rotation, 3-year, 6-year, and 20-year mugwort cropping) on soil microbial community in temperate regions of Northern China. The results showed that the highest total, actinomycetes, and fungi PLFAs were found in the 3-year mugwort cropping soils. All PLFAs of microbial groups were lowest in the 20-year mugwort cropping soils. All of the three cropping years of mugwort increased network complexity of soil microbial community. Changes in total nitrogen and phosphorus content as well as the ratio of ammonium nitrogen to nitrate nitrogen could be primarily explain the variations in soil microbial community along the mugwort cropping chronosequence. Our observations highlight the contrasting impacts of soil microbial community to short-term and long-term mugwort cropping compared to conventional rotations and would have critical implications for sustainable agricultural management under perennial cropping in temperate regions.

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.

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.

Eating and drinking difficulties are common in dementia, but little is known about the experiences of ethnic minority groups managing these difficulties at home. We undertook qualitative semi-structured interviews, exploring the meaning of food, the impact of dementia on eating and drinking and carers’ experiences of support. Interviews were audio-recorded and transcribed verbatim. We interviewed 17 carers and people with dementia from ethnic minority backgrounds, using reflexive thematic analysis to analyse data. Food/drink had strong links to identity, culture and emotions. Providing culturally familiar foods, celebrating traditional festivals and supporting previous food-related roles promoted reminiscence, which encouraged people with dementia to eat and drink, as did social interactions. However, these strategies sometimes led to distress in those with more advanced dementia. Food choices were also influenced by carer strain, generational differences and the impact of health conditions. Despite a strong sense of duty to care for relatives at home, there was low awareness of community support services. Carers expressed a need for culturally tailored support for managing dementia-related eating and drinking difficulties at home. Healthcare professionals must provide contextually relevant advice to carers, being mindful of how cultural backgrounds can affect dietary choices.

The impacts of climate change on shrimp aquaculture can vary widely and can have environmental and socioeconomic consequences. This study assessed the vulnerability to climate change impacts of selected small-scale shrimp farms of Penaeus vannamei and shrimpfish market vendors in Davao region, Philippines using a modified Fisheries Vulnerability Assessment Tool (FishVool). Shrimp farmers and vendors were interviewed using two separate semi-structured questionnaires. A total of thirty-nine (N=39) shrimp farmers and forty-eight (N=48) market vendors from various market areas within the region were interviewed. Data regarding exposure (E), sensitivity (S), and adaptive capacity (AC) were collected following the FishVool parameters with modifications. Results revealed that overall climate change vulnerability of the shrimp farmers was medium (M), where both exposure and adaptive capacity were low (L) while sensitivity was medium (M). In addition, the shrimp market vulnerability of the various sites examined revealed medium (M) scores for markets in Pantukan, Mabini, Tagum, Maco, Lupon, Davao City, and Digos. But high (H) vulnerability scores for the markets in Panabo and Sta Cruz. Overall, the study provided a better understanding about shrimp farming in relation to climate change impacts and vulnerability and provided information for future shrimp farm management, marketing and climate change adapation in the region.

Cultural recovery of Mucorales from hyphae-laden tissue is poor, and the clinical implications of culture positivity are scarcely studied. Therefore, we compared clinical and histological characteristics of culture-positive and culture-negative histology-proven pulmonary mucormycosis cases among cancer patients. Histology specimens were blindly reviewed by a Thoracic Pathologist and graded on four histopathologic features: hyphal quantity, tissue necrosis, tissue invasion, and vascular invasion. Twenty cases with a corresponding fungal culture were identified; five were culture-positive, and 15 culture-negative. Although no statistically significant differences were found, culture-positive patients were more likely to exhibit a high burden of necrosis and have a high burden of hyphae but tended to have less vascular invasion than culture-negative patients. In terms of clinical characteristics, culture-positive patients were more likely to have acute myeloid leukemia (60% vs. 27%, p=0.19), a history of hematopoietic cell transplant (80% vs. 53%, p=0.31), severe lymphopenia (absolute lymphocyte count ≤500/µL, 100% vs. 73%, p=0.36), and monocytopenia (absolute monocyte count ≤100/µL, 60% vs. 20%, p=0.11). Forty-two-day all-cause mortality was comparable between culture-positive and culture-negative patients (60% and 53%, p=0.80). This pilot study represents the first comprehensive histopathological scoring method to examine the relationship between histopathologic features, culture positivity, and clinical features of pulmonary mucormycosis.

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.

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.

Cyanobacteria are extensively studied and cultured because they can produce many value-added substances among which are pigments, mainly the phycobiliproteins phycocyanin (PC), phycoerythrin (PE), allophycocyanin (APC) and chlorophyll-a and carotenoids as well. As numerous cyanobacterial species await optimization for maximizing pigment production, we examined here two local marine species, Phormidium sp. and Cyanothece sp. batch cultured under 18-19.5 oC, at 40 ppt salinity with Walne’s nutrient medium, using white LED light of low (2000 lux) and high (8000 lux) intensity and additionally blue, green and red LED light. Significant differences were found among the intensities and colors of light used. Maximum growth was induced by high white light in both species (2.15 g dw/L in Phormidium and 1.47 g/L in Cyanothece). Next to them was green light (1.25 g/L) in Cyanothece and low white and green (1.26 – 1.33 g/L) in Phormidium. Green light maximized phycocyanin content in Phormidium (0.45 mg/mL), while phycoerythrin was maximized (0.17 mg/mL) by blue light and allophycocyanin by all colors (~0.80 mg/mL). All colors maximized phycocyanin in Cyanothece (~0.32 mg/mL) while phycoerythrin and allophycocyanin were maximized under green light (~0.138 and 0.38 mg/mL respectively). In Phormidium maximization of chlorophyll-a (9.3 μg/mL) was induced by green light while total carotenoids and b-carotene (3.05 and 0.89 μg/mL respectively) by high white light. In Cyanothece both white light intensities along with green light maximized chlorophyll-a content (~9 μg/mL) while high white light and green maximized total carotenoids (2.6-3.0 μg/mL).

Phenotypic plasticity in response to adverse conditions determines plant productivity and survival. The aim of this study was to test if two highly productive Populus genotypes, characterized by different in vitro etiolation patterns, differ also in their responses to hormones gibberellin (GA) and abscisic acid (ABA), and to a GA biosynthesis inhibitor paclobutrazol (PBZ). The experiments on shoot cultures of ‘Hybrida 275’ (abbr. H275; Populus maximowiczii × P. trichocarpa) and IBL 91/78 (Populus tremula × P. alba) were conducted either by modulating the physical in vitro environment or by adding specific chemicals to the nutrient medium. Our results show that there are significant differences between the studied genotypes in environmental and hormonal regulation of growth responses. The genotype H275, which responded to darkness with PBZ-inhibitable shoot elongation, was unable to recover its growth after treatment with ABA. In contrast, the genotype IBL 91/78, whose shoot elongation was not affected either by darkness or PBZ treatment, recovered so well after the ABA treatment that, when rooted subsequently, it developed longer shoots and roots than without ABA treatment. Our results indicate that GA catabolism and repressive signaling provide an important pathway to control growth and physiological adaptation in response to immediate or impending adverse conditions. These observations can help breeders define robust criteria for identifying genotypes with high resistance and productivity and highlight where genotypes exhibit susceptibility to stress.

The process of full-thickness skin regeneration is complex and has many parameters involved, which makes it difficult to use a single dressing to meet the various requirements of the complete regeneration at the same time. Therefore, developing hydrogel dressings with multifunction, including tunable rheological properties and aperture, hemostatic, antibacterial and super cytocompatibility, is a desirable candidate in wound healing. In this study, a series of complex hydrogels were developed via the hydrogen bond and covalent bond between chitosan (CS) and alginate (SA). These hydrogels exhibited suitable pore size and tunable rheological properties for cell adhesion. Chitosan endowed hemostatic, antibacterial properties and great cytocompatibility and thus solved two primary problems in the early stage of the wound healing process. Moreover, the sustained cytocompatibility of the hydrogels was further investigated after adding FGF and VE-cadherin via the co-culture of L929 and EC for 12 days. The confocal 3D fluorescent images showed that the cells were spherical and tended to form multicellular spheroids, which distributed in about 40-60μm thick hydrogels. Furthermore, the hydrogel dressings significantly accelerate defected skin turn to normal skin with proper epithelial thickness and new blood vessels and hair follicles through the histological analysis of in vivo wound healing. The findings mentioned above demonstrated that the CS/SA hydrogels with growth factors have tremendous potential as multifunctional hydrogel dressings for full-thickness skin regeneration incorporated with hemostatic, antibacterial, sustained cytocompatibility for 3D cell culture and normal skin repairing.

Museums increasingly recognize the need to address advances in digital culture which impact the expectations and needs of their audiences. Museum collections of real objects need to be presented both on their own premises and digitally online, especially as social media becomes more and more influential in people’s everyday lives. We investigate these challenges magnified by advances in digital and computational media and culture looking particularly at recent and relevant reports on changes in the ways museums interact with the public. We find that the Covid-19 pandemic has accelerated many of the changes driving museum transformation. We believe that museums must be more prepared than ever to adapt to unabated technological advances set in the midst of cultural and social revolution, now intrinsic to the digital landscape in which museums are inevitably connected and participating across the global digital ecosystem where they inevitably find themselves entrenched.

The production of vaccines of biological origin presents a tremendous challenge for researchers. In this context, animal cell cultures are an excellent alternative for the isolation and production of biologicals against several viruses since they have an affinity with viruses and a great capacity for their replicability. Different variables have been studied to know the system's ideal parameters, allowing it to obtain profitable and competitive products. Consequently, this work focuses its efforts on evaluating an alternative for producing an anti-influenza biological from MDCK cells using SuperPro Designer v8.0 software. The process uses the DMEN culture medium supplemented with nutrients as raw material for cell development; the MDCK cells were obtained from a potential scale-up with a final working volume of 500 L, four days of residence time, inoculum volume of 10%, and continuous working mode with up to a total of 7400 h/Yr of work. The scheme has the necessary equipment for the vaccine's production, infection, and manufacture with yields of up to 416,698 units/h. In addition, it was estimated to be economically viable to produce recombinant vaccines with competitive prices of up to 0.31 USD/unit.

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.

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.

Hormones must be balanced and dynamically controlled for the Female Reproductive Tract (FRT) to function correctly during the menstrual cycle, pregnancy, and delivery. Gamete selection and successful transfer to the uterus, where it implants and pregnancy occurs, is supported by the mucosal epithelial lining of the FRT ovaries, uterus, cervix, fallopian tubes, and vagina. Successful implantation and placentation in humans and other animals rely on complex interactions between the embryo and a receptive female reproductive system. The FRT's recent breakthroughs in three-dimensional (3D) organoid systems now provide critical experimental models that match the organ's physiological, functional, and anatomical characteristics in vitro. This article summarizes the current state of the art on organoids generated from various parts of the FRT. The current analysis examines recent developments in the creation of organoid models of reproductive organs, as well as their future directions.

Cyanobacteria are microorganisms able to adapt to a wide variety of environmental conditions and abiotic stresses. They produce a very large number of metabolites that can participate in the adaptation of cyanobacteria to a large range of resources such as light, temperature, or nutrient. The metabolites variation is one way to understand the physiological status and adaptation of cells. In this study, we aim to understand how the diversity and the dynamics of the whole metabolome is dependent of the growth phases and under control of abiotic factors (e.g. light intensity and temperature). The cyanobacteria Aliinostoc sp. PMC 882.14 was selected for its large number of biosynthetic gene clusters. Metabolomes were analyzed by using mass spectrometry (qTOF-MS/MS) combined with untargeted analysis to investigate the metabolite dynamics. Significant variations were characterized between exponential and stationary phases, whatever the culture conditions (“control”, “higher light”, or “higher temperature”). ”Higher light” and “higher temperature” favored the synthesis of metabolites belonging to the same molecular families. Among highly regulated metabolites, we observe the presence of mycosporine-like amino acids (MAAs), and various variants of somamides, microginins, and microviridins. Through Aliinostoc sp. PMC 882.14, this study shows the importance of knowing the physiological state of cyanobacteria for comparative global metabolomics and questions the regulation processes involve into metabolite families production. Our results also open up new perspectives in the context of the production of targeted bioactive metabolites.

Alzheimer’s disease (AD) is a significant health concern worldwide with enormous social and economic impact globally. The gradual deterioration of cognitive functions and irreversible neuronal losses are primary features of the disease. Even after decades of research, most therapeutic options are merely symptomatic, and drugs in clinical practice present numerous side effects. Lack of effective diagnostic techniques prevents the early prognosis of disease, resulting in a gradual deterioration in the quality of life. Furthermore, the mechanism of cognitive impairment and AD pathophysiology is poorly understood. Microfluidics exploits different microscale properties of fluids to mimic environments on microfluidic chip-like devices. These miniature multichambered devices can be used to grow cells and 3D tissues in vitro, analyze cell-to-cell communication, decipher the roles of neural cells like microglia, and gain insights into AD pathophysiology. This review focuses on the applications and impact of microfluidics on AD research. We discuss the technical challenges and possible solutions provided by this new cutting-edge technique to understand disease-associated pathways and mechanisms.

An increasing number of studies have shown that safety culture factors have a substantial influence on safety performance in a variety of industrial sectors. These factors' impact on safety performance is unclear, especially at the public service and statutory authorities. On the other hand, the understanding of indicators for safety performance in every working sector in Malaysia is on the continuing progress. Hence, this study's contribution is to explore the influence of safety culture factors (i.e., management commitment and supervision in safety, safety system) and safety competence on safety performance in government paramedic training institutes. IPMA (importance-performance map analysis) is a technique used in Smart PLS to determine the significance and performance of each of these factors. The study was conducted via an online survey and involved 258 safety and health committee members in Ministry of Health paramedic training institute. As a matter of relevance, the IPMA's empirical data study revealed that management commitment and supervision in safety were the predominant factors in determining safety performance. Meanwhile, for performance, the findings showed that worker involvement, safety system, and safety competence perform well in determining the safety performance.

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.

Nowadays, probiotics has been proposed for substituting antibiotics in animal feed, since the European Union (EU) banned the latter compounds in 2006 to avoid their serious side effects on human health. Therefore, this work aimed to produce a probiotic product by fed-fermentation of whey with kefir grains for use in animal feed. The whey was fermented with a combination of kefir grains AGK1 and the free biomass present in whole milk fermented milk used to activate these kefir grains. The probiotic culture obtained was characterized with high levels of biomass, total viability and antibacterial activity. Some probiotic properties of the probiotic culture were investigated in vitro, including its survival at low pH values, under simulated gastrointestinal conditions, after freezing in skim milk at − 20 ºC, and in the commercial feed during storage at room temperature. The viable cells of lactic and acetic acid bacteria and yeasts exhibited higher tolerance to acidic pH and simulated gastrointestinal conditions when the cells were protected with skim milk and piglet feed, compared with washed cells. The results indicated the feasibility of producing a probiotic product at a low cost with a potential application in animal feed.

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.

Malassezia species are fastidious and slow-growing yeasts whose isolation from polymicrobial samples is hampered by fast-growing microorganisms. Malassezia selective culture media are needed because Malassezia are resistant to cycloheximide, but some fungi, including the chief human commensal Candida albicans resist to this compound. This study aimed to test whether the macrolide rapamycin could be used in combination with cycloheximide to develop a Malassezia-selective culture medium. Rapamycin susceptibility testing was performed via microdilution assays in modified Dixon against M. furfur and five Candida spp. The MIC was the lowest concentration producing 90% growth inhibition. Rapamycin medium ± cycloheximide 500 mg/L was also added to FastFung solid and yeast suspensions were inoculated and incubated for 72h. Rapamycin MICs against Candida spp. ranged from 0.5 to 2 mg/L, except for C. krusei whose MIC was >32 mg/L. M. furfur stains were rapamycin resistant. Rapamycin and cycloheximide supplementation of the FastFung medium effectively inhibited the growth of non-Malassezia yeast, including the cycloheximide-resistant C. albicans and C. tropicalis. Based on our findings, we recommend using this “MalaSelect” medium for Malassezia isolation and culture from polymicrobial samples.

A strain of the filamentous non N-fixing cyanobacterium Phormidium sp. isolated from the Messolonghi (W. Greece) saltworks, was cultured in the laboratory at 6 different combinations of salinity (20-40-60 ppt) and illumination (low-2000 lux and high-8000 lux). At salinities of 60 and 40 ppt and in high illumination (XL-8000 lux) the growth rate (μmax) presented the highest values (0.491 and 0.401 respectively) compared to the corresponding at 20 ppt (0.203). In general and at all salinities, the higher illumination (XL) gave the highest growth rates and shorter dublication time (tg) in comparison to the lower illumination (L). On the contrary, phycocyanin, phycoerythrin and allophycocyanin production was extremely increased in the lower illumination (L) in all salinities, from ~14fold at 40 and 60 ppt to 269fold at 20 ppt of those corresponding to higher illumination (XL). Similar analogies were also recorded for the other two billiproteins. Chlorophyll-a content was also higher in lower illumination at all salinities in contrast to total carotenoids that did not exhibit such a pattern. The high growth rate and high phycocyanin content along with the rapid sedimentation of its cultured biomass can set this marine Phormidium species as a promising canditate for mass culture.

Cannabis has developed into a multi-billion dollar industry that relies on clonal propagation of elite genetics with desirable agronomic and chemical phenotypes. While the goal of clonal propagation is to produce genetically uniform plants, somatic mutations can accumulate during growth and compromise long-term genetic fidelity. Cryopreservation is a process in which tissues are stored at cryogenic temperatures, halting cell division and metabolic processes to facilitate high fidelity germplasm preservation. In this study, a series of experiments were conducted to optimize various stages of cryopreservation and develop a protocol for long-term germplasm storage of Cannabis sativa. The resulting protocol uses a standard vitrification procedure to cryopreserve nodal explants from in vitro shoots as follows: Nodes were cultured for 17 hours in a pre-culture solution (PCS), followed by a 20 minute treatment in a loading solution (LS), and a 60 minute incubation in plant vitrification solution 2 (PVS2). The nodes were then flash frozen in liquid nitrogen, re-warmed in an unloading solution at 40°C, and cultured on basal MS culture medium in the dark for 5 days followed by transfer to standard culture conditions. This protocol was tested across 13 genotypes to assess the genotypic variability. The protocol was successful across all 13 genotypes, but significant variation was observed in tissue survival (43.3-80%) and regrowth of shoots (26.7-66.7%). Plants grown from cryopreserved samples were morphologically and chemically similar to control plants for most major traits, but some differences were observed in the minor cannabinoid and terpene profiles. While further improvements are likely possible, this study provides a functional cryopreservation system that works across multiple commercial genotypes for long-term germplasm preservation.

As many as 20-25% of the population experiences harm in outpatient settings, yet these locations are underrepresented in the literature compared to hospitals. We examined results from the Medical Office Survey on Patient Safety Culture designed by the Agency for Healthcare Research and Quality. The survey administered in 2012 gathered perceptions from 23,679 individuals in 934 unique medical offices. We examined associations of organizational patient safety climate composites on frequency of safety and quality issues, overall quality score, and safety rating. We found organizational patient safety composites are all positively and significantly associated with a higher overall quality score and patient safety rating, and fewer safety and quality issues. Office processes and standardization appeared to have the most consistent influence on perceived quality outcomes. Our results indicate it may be advantageous for medical offices to improve on the factors that contribute to positive safety climate.

Philip Phenix’s (1964) book Realms of meaning started the ever growing movement concerned with how school education might help young people in their search for meaning in times of rapid social change. Today, in globalised, digital, secularised, de-traditionalised culture, the importance and urgency of this role have never been greater. Cultural change has accelerated exponentially, and for many – including students in religious/Catholic schools – traditional religious sources of meaning are no longer prominent or plausible reference points. Catholic schools, whether independent or semi-state institutions because of government funding, can make a valuable contribution young people’s spiritual/moral education, no matter what their level of religious affiliation or practice. This article argues that such a contribution requires change to the discourse or narrative of Catholic school Religious Education, with corresponding adjustments to content and pedagogy. Its present trajectory, which is excessively concerned with promoting a Catholic identity in students, needs to be modified. Both the religious and non-religious students, especially in the senior classes, would derive greater spiritual and religious benefit from the inclusion of more life-relevant and issue-related content, together with a critical, research-oriented pedagogy. Such an approach proposes that the Catholic Church’s schools should offer unconditionally a meaningful spiritual/moral education that is relevant to all students, rather than a traditional one which seemed to presume that all students are, or should be practising Catholics. This does not minimise attention to the Catholic tradition, but it allows for a study of how people negotiate the task of constructing meaning and values in a complex culture. The article also looks at the ‘headwinds’ that hinder the implementation of this approach. The article is focused specifically on the Australian context where Catholic schools are semi-state institutions because they are funded by both state and federal governments. The issues are still likely to be pertinent to Catholic education in other countries, while taking into account significant contextual differences.

Background: The saga of repatriating cultural artefacts continues as western museums face increasing pressure from claimants. Western museums that have been involved in the display of historical artefacts, most of which were acquired during the colonial period, have come under huge criticism. A heated discussion of late has been the legitimacy of retaining artefacts in western museums. This study aimed at investigating the ongoing debate regarding the restitution of artefacts. Objective: To investigate the arguments for and against the repatriation of artefacts in relation to diplomatic exchange, preservation, legitimacy and usefulness. Methods: Records will be searched in electronic databases including the University of Manchester Library for Social Anthropology, Scorpus and Project MUSE. Search terms will include "return of artefacts", “return of historical objects”, “return of cultural objects”, “western museums”, “restitution of artefacts”, “repatriation of artefacts”, “restitution of historical objects”, repatriation of historical objects”, “restitution of cultural objects”, “repatriation of cultural objects”, "material culture", "return of antiquities”, restitution of antiquities” and “repatriation of antiquities”. Coding and analysis will be done in SWIFT-Review. The deductive and inductive approaches will be used in synthesising results. Both tabular and graphical methods will be used to present results. Ethics and Results: This study did not need any ethical approval. Results on study characteristics, quality and risk of bias assessments as well as the synthesis of arguments for and against the restitution of artefacts will be presented. The review results will be reported according to appropriate guidelines and disseminated through publication in a relevant journal and presented to stakeholders where necessary. Conclusions: This review will be based on current protocols for systematic review and qualitative evidence synthesis. The study will be the first review that seeks to pull together claims for and against the return of cultural artefacts. The conclusions that will be drawn and recommendations will provide the basis for further research into the debate and the way forward. This study will also help identify the existing gaps regarding the subject matter.

In modern times like today, technology and science are developing very fast. This has resulted in a condition known as globalization. This era has had a significant impact on every country globally, including in Indonesia, such as the spread of culture is solid. This flow has many benefits and advantages. However, this globalization can also have a negative impact. One of them is the erosion of native Indonesian culture compared to foreign cultures. Therefore, this research to find the origin, causes, and impacts of the influx of cultural globalization on the current culture of East Java because it is the province of origin of the researchers. Here the researchers asks about the impact of this modern culture and how influential culture is to the people of East Java. The method used by the researchers in making this paper is Literature Review using qualitative descriptive writing with quantitative data. The researchers findings as material in this paper amounted to 24 materials with the provisions of 20 journal article materials, 2 article materials online provided that they were published in the last three years, and 2 E-books for methods on this paper. This study's boundaries are a modern culture that the researchers takes about western culture (western) and South Korean culture (Hallyu). In addition, this study only discusses cultures originating in East Java, such as Javanese.

(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.

Ethylene is a plant hormone controlling physiological and developmental processes such fruit maturation, hairy root formation and leaf abscission. Its effect on regeneration systems, such as organogenesis and somatic embryogenesis (SE), has been studied and progresses in molecular biology techniques have contributed to unveil mechanisms behind its effects. This compound affects regeneration differently, depending on the species, genotype and explant. In some species, ethylene seems to revert recalcitrance in genotypes with low regeneration capacity. However, its effect is not addictive, since in genotypes with high regeneration capacity this ability decreases in the presence of ethylene precursors, suggesting that regeneration is modulated by ethylene. Several lines of evidence have shown that the role of ethylene on regeneration is markedly connected to biotic and abiotic stresses as well as to hormonal-crosstalk, in particular with key regeneration hormones and growth regulators of the auxin and cytokinin families. Transcriptional factors of the ethylene response factor (ERF) family are regulated by ethylene and strongly connected to SE induction. Thus, an evident connection between ethylene, stress responses and regeneration capacity is markedly established. In this review the effect of ethylene and the way it interacts with other players during organogenesis and somatic embryogenesis is discussed.

We explore gaps in Wikipedia's coverage of the visual arts by comparing the representation of 100 artists and 100 artworks from the Western canon against corresponding sets of notable artists and artworks from non-Western cultures. We measure the coverage of these two sets of topics across Wikipedia as a whole and for its individual language versions. We also compare the coverage for Wikimedia Commons and Wikidata, sister-projects of Wikipedia that host digital media and structured data. We show that all these platforms strongly favour the Western canon, giving many times more coverage to Western art. We highlight specific examples of differing coverage of visual art inside and outside the Western canon. We find that European language versions of Wikipedia are generally more "Western" in their coverage and Asian languages more "global", with interesting exceptions. We suggest how both Wikipedia and the wider cultural sector can address this gap in content and thus give Wikipedia a truly global perspective on the visual arts.

Background: and objectives: Covid-19 is a global pandemic that requires a culturally integrated response in all Latin American government systems. In this study, we investigate the multidimensional associations of actions in Latin American indigenous peoples against Covid-19 Methods: We performed a data mining analysis of scientific literature records from 19 academic journals, using the topics Covid-19, mortality, indigenous peoples, and future measures, rights and actions. The literature is analyzed in a multidimensional way, either isolated or combined. It uses switchable statistical methods, the R Studio, worddj, Gephi, and Iramuteq for both textual and multidimensional analysis. Results and Conclusions: In this analysis, the literature is classified into 4 linked groups which are the indigenous people, the rights of territorial protection, the indigenous health culture, the Covid-19 pandemic with its protection efforts. Taking the indigenous peoples as an axis, we observe that the covid pandemic and the protection of their territories do not have the same force in the current context, the indigenous people are more linked to the protection of their territory. Indigenous peoples have a link of cultural strength in traditional health independent of Covid treatment. It should be noted that government agencies are taking measures regarding the covid pandemic with an effort not marked within the health culture of indigenous peoples.

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.

Candida auris is an emerging multidrug resistant yeast causing nosocomial infections and associated with high mortality in immunocompromised patients. Rapid identification and characterisation is necessary for its diagnosis and containing spread. In this study, we present a selective culture medium for all C. auris clades. This medium is sensitive with a limit of detection of 102 CFU/ml. The 100% specificity of SCA (Specific C. auris) medium is confirmed on a set of 134 Candida strains, 50 bacterial species and 200 human stool samples. Thus, this medium specifically selects for C. auris isolation from clinical samples, and allows studying its phenotypic profile.

Background: Circulating tumor cells (CTC) have relevance as prognostic markers in breast cancer. However, the functional properties of CTCs or their molecular characterization have not been well-studied. Experimental models indicate that only a few cells can survive in the circulation and eventually metastasize. Thus, it is essential to identify these surviving cells capable of forming such metastases. Methods: We isolated viable CTCs from 50 peripheral blood samples obtained from 35 patients with advanced metastatic breast cancer using RosetteSepTM for ex vivo culture. The CTCs were seeded and monitored on plates under low adherence conditions and with media supplemented with growth factors and Nanoemulsions. Phenotypic analysis was performed by immunofluorescence and gene expression analysis using RT-PCR and CTCs counting by Cellsearch® system. Results: We found that in 75% of samples the CTC cultures lasted more than 23 days, predicting a shorter Progression-Free Survival in these patients, independently of having ≥ 5 CTC by Cellsearch®. We also observed that CTCs before and after culture showed a different gene expression profile. Conclusions: the cultivability of CTCs is a predictive factor. Furthermore, the subset of cells capable of growing ex vivo show stem or mesenchymal features and may represent the CTC population with metastatic potential in vivo.

Clinical education is a method that is applied to formal nurse education as a step to provide real and direct learning experiences in the nursing environment correctly and effectively. The success of education in a clinical setting certainly requires the support of teaching nurses (clinical instructors) who have credibility and competence in terms of knowledge, attitudes and skills and are actively involved in professional activities. The diversity of backgrounds of nurses and students, including patients, certainly contributes to a shift in paradigms and perspectives for the nursing environment both in education and in clinical settings in health services. Responding to this cultural diversity, it is important to prepare knowledge and understanding related to transcultural nursing issues, intercultural communication and clinical education which explores the socio-cultural elements in the implementation of staff, students and patients. Purpose: The purpose of this literature review is to identify the extent to which nurse educators play a role by including socio-cultural and transcultural aspects in efforts to develop the quality of education in clinical practice environments in the millennium era. Method: The method of writing this article uses 11 literature review, the publication year period 2019,2020 and 2021 with sources from 4 databases such as science Direct, Scopus, ProQues and Elsevier. The review guidelines used are based on Prisma and the Joanna Briggs Institutute. The level of eligibility is identified through the title, abstract, research methodology as well as the type of scholarly journal and full text. Results: The results of the reviews found are presented in a narrative form. The results of the review study found that there were 11 articles explaining the competence of clinical education based on the socio-cultural approach, which is an educational strategy in the clinical area that integrates transcultural elements of nursing, intercultural communication, collaboration, self-directed with the principles of openness, honesty, and mutual respect in the implementation of team interaction and collaboration. The development of interpersonal relationships is also an important role that educators must have in helping to introduce the nurse orientation process to the organizational environment and other professional teams so that the achievement of satisfaction with clinical education is able to improve the performance of nurses and students perfectly. Conclusion: Clinical education which is supported by the competence of nurse educators (clinical instructors) who have individual and professional competences has a role to play in improving clinical learning outcomes by both students and nurses with a socio-cultural and transcultural strategic approach that will create satisfaction with the achievement of clinical competence and performance effectively.

The number of entrepreneurship education programs (EEP) has increased exponentially over the past two decades. However, a systematic review has not yet been carried out to confirm the effectiveness of EEPs and their presence in the current global world. The main objective of this study was to provide a systematic synthesis of EEP, exploring their characteristics and effectiveness. The search was carried out in the following databases: Scopus, Web of Science, ProQuest and ERIC. Twenty-nine articles were included, with programs developed mainly in European (n=15), Asian (n=6) and American (n=5) countries. The programs were mainly aimed at higher education students (n=17), addressing the development of business plans and entrepreneurial skills. However, greater attention is paid to the entrepreneurial skills in both basic and secondary education. The development of the programs under analysis varied between one week and two years. The results of the studies showed the effectiveness of most of these programs in promoting entrepreneurial skills at all levels of education. In turn, there was not a verified increase in the intention to start a business, since this intention is determined by predisposition, namely socio-cultural and family aspects. This systematic review of the EEP points to the need for this type of program to be preferentially developed since the early school years, since it is at that time that predispositions are created for the development of entrepreneurial skills and intentions. This condition is corroborated by the global geography of the EEP, which demonstrates that, where there is currently an entrepreneurial culture, countries have made a long educational journey, with strategic options from the perspective of educational policies defending entrepreneurship among the younger generations.

There is a growing consensus that culture influences the perception of facial expressions of emotion. However, little is known about whether and how culture shapes the production of emotional facial expressions, and even less so about whether culture differentially shapes the production of posed versus spontaneous expressions. Drawing on prior work on cultural differences in emotional communication, we tested the prediction that people from the Netherlands (a historically heterogeneous culture where people are prone to low-context communication) produce facial expressions that are more distinct across emotions compared to people from China (a historically homogeneous culture where people are prone to high-context communication). Furthermore, we examined whether the degree of distinctiveness varies across posed and spontaneous expressions. Dutch and Chinese participants were instructed to either pose facial expressions of anger and disgust, or to share autobiographical events that elicited spontaneous expressions of anger or disgust. Using the complementary approaches of supervised machine learning and information-theoretic analysis of facial muscle movements, we show that posed and spontaneous facial expressions of anger and disgust were more distinct when produced by Dutch compared to Chinese participants. These findings shed new light on the role of culture in emotional communication by demonstrating, for the first time, effects on the distinctiveness of production of facial expressions.

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.

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.

Abstract. Background: This paper investigates the propagation of behaviorally transmitted traits with negative effect on host fitness. Methods: We analyze equilibrium between genetically transmitted and behaviorally transmitted competing propagators and consider whether a behavioral propagator is linked to reproduction (e.g. vertical culture transmission), or not. We employ combined genetic and behavior-induced fitness components for hosts, while behavioral propagators have replication factors to distinguish from what’s good for the host (fitness). Results: A trait which spreads faster than its marginal host fitness contribution reduces population will establish itself. The often transient nature of laterally transmitted traits may be a defense against accumulation of deleterious traits. Laterally transmitted traits with high spreading rate often do not equalize with genetic traits, spreading outside natural selection of the hosts. Vertical transmission reduces replication rate and allows group selection against deleterious behaviorally transmitted traits. Competing mutually exclusive propagators contribute to inequality and altruism, but compete through adverse fitness since exclusivity assumes low conversion. Conclusion: Behaviorally transmitted traits, in some cases a tremendous advantage, may also be a significant problem in the development of societies.

Internal and external factors influence the tendency of corruption. One of these internal factors is Dark Triad Personality, religiosity, and compensation, while the external factors are the organizational culture. This first study was conducted on 222 private employees and the second study 205 state employee in the East Java area. All subjects have worked at the company for at least 1 year and have a position as the staff, administrator, manager, and age range from 22 to 55. The results of the first study using regression analysis showed that: there was a significant correlation between dark triad personality, organization culture, and the tendency of corruption of private employee (F = 60,132; p = 0,000); there was a significant negative correlation between organizational culture and tendency of corruption (p = 0,000); there was a positive correlation between dark triad personality and the tendency of corruption (p = 0.05). The results of the second study using regression analysis showed that: there was a significant correlation between religiosity, compensation, and the tendency of corruption of state employee (F = 65.950; p = 0,000); there was a significant negative correlation between religiosity and tendency of corruption (p = 0,000); there was no correlation between compensation and tendency of corruption (p = 0.074). This research's practical implication is: the company or institution must always create a positive perception of organizational culture, minimize dark triad personality, and maximize religiosity to reduce the tendency of corruption among employees.

The recent legalization of Cannabis sativa L. in many regions has revealed a need for effective propagation and biotechnologies for the species. Micropropagation affords researchers and producers methods to rapidly propagate insect/disease/virus free clonal plants, store germplasm, and forms the basis for other biotechnologies. Despite this need, research in the area is limited due to the long history or prohibitions and restrictions. Existing literature has multiple limitation: many publications use hemp as a proxy for drug-type Cannabis when it is well established that there is significant genotype specificity, studies using drug-type cultivars are predominantly op-timized using a single cultivar, most protocols have not been replicated by independent groups, and some attempts demonstrate a lack of reproducibility across genotypes. Due to culture decline and other problems the multiplication phase of micropropagation (stage 2) has not been fully developed in many reports. This review will provide a brief background on the history and botany of Cannabis as well as a comprehensive and critical summary of Cannabis tissue culture. Special attention will be paid to current challenges faced by researchers, the limitations of existing Cannabis micropropagation studies, and recent developments and future directions of Cannabis tissue culture technologies.

The mutualistic interactions between the oleaginous yeast Lipomyces starkeyi and the green mi-croalga Chloroidium saccharophilum in mixed cultures were investigated to exploit possible syner-gistic effects. As a matter of facts, microalga could act as an oxygen generator for the yeast, while the yeast could provide carbon dioxide to microalga. A lignocellulosic hydrolysate from steam exploded Arundo donax (Giant reed) was used as low cost feedstock. The overall lipid content and lipid productivity obtained in the mixed culture treating the hydrolysate of Arundo donax were equal to 0.081 glipid.gbiomass-1 and 37.2 mglipid.L-1.d-1, respectively. They represented promising re-sults if compared to the model systems where synthetic media were used. This study provided new input for the integration of Single Cell Oil (SCO) production with agro-industrial feedstock and the fatty acid distribution mainly consisting of stearic (C18:0) and oleic acid (C18:1) allows promising applications in biofuels, cosmetics, food additives and other products of industrial interest.

As a result of the influence of geographical environment and historical heritage, food preference has significant regional differentiation characteristics. However, the spatial structure of food culture represented by the cuisine culture at the regional level has not yet been explored from the perspective of geography. This study aims to explore such patterns by focusing on the restaurants of the eight most famous cuisines in Mainland China. Initially, the density based geospatial hotspot detector method is proposed to analyze and mapping the spatial quantitative characteristics of the eight major cuisines. A heuristic method for geographical regionalization based on machine learning was used to analyze spatial distribution patterns in accordance with the proportion of these cuisines in each prefecture-level city. Results show that some types of single-category cuisines have a stronger spatial concentration effect in the present, whereas others have a strong diffusion trend. In the comprehensive analysis of multicategory cuisines, the eight major cuisines formed a new structure of geographical regionalization of Chinese cuisine culture. This study is helpful to understand regional structure characteristics of food preference, and the density based hotspot detector proposed in this paper can also be used in the analysis of other type of POI data.

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.

Economic sustainability of rural areas is a concern given the increasing trend towards urbanization globally and in India. Self-reliant strategies, including increased savings and investment are more valuable in this regard than external interventions. This paper uses the results of a survey to examine the factors affecting saving in a rural part of Odisha populated primarily by tribals. Our tentative findings are that savings propensity is determined partly by the extent to which individuals feel connected to the broader economy, and partly by cultural factors. One implication of these findings is that connecting rural areas to other, possibly urban, locations could elicit greater saving and this could lead to greater development, employment possibilities, economic betterment and all the consequent social welfare implications. This paper relates savings propensity to new sociological population characteristics, such as perceived connectivity and food consumption patterns, and hence provides hitherto unexplored clues for policy initiatives to increase savings.

Considered one of the finest first-generation Nigerian writers, John Pepper Clark-Bekederemo, who passed away on 13th October, 2020, had been categorised as a Eurocentric writer. This work assesses the authenticity or otherwise of this critics’ perception of Clark by critiquing his America, their America. By analysing this autobiography vis-à-vis the notion of self and other which is a theoretical concern in contemporary travel writing, it was established, among other things, that every culture has its dark sides which it must not feel too proud to change as time and situation demand; that Clark vehemently rejects the Americans’ claim of sophistication and superiority of their culture over African culture. The paper concludes that contemporary travel writing should be a rightful site for negotiating cultural, political and diplomatic compromises between the Self and the Other since the gulf may be difficult to close altogether.

Eysenhardtia platycarpa (Fabaceae) is a medicinal plant used in México and it lacks biotechnological studies for its use. The aim of this work was to establish a cell suspension cultures (CSC) of E. platycarpa, determine the phytochemical profile, and evaluate its antifungal activity. Friable callus and CSC were established with 2 mg/L 1-naphthaleneacetic acid plus 0.1 mg/L kinetin. The highest total phenolics of CSC was 15.6 mg GAE/g dry weight and the total flavonoids content ranged from 56.2 to 104.1 µg QE/g dry weight. CG‒MS analysis showed that the dichloromethane extracts of CSC, sapwood and heartwood have a high amount of hexadecanoic acid (22.3 ‒ 35.3 %) and steroids (13.5 ‒ 14.7%). Heartwood and sapwood defatted hexane extracts have the highest amount of stigmasterol (≈ 23.4%) and β-sitosterol (≈ 43%), and leaf extracts presented β-amyrin (16.3%). Methanolic leaves extracts showed mostly sugars and some polyols, mainly D-pinitol (74.3%). Dichloromethane and fatty hexane extracts of CSC exhibited the percentages inhibition higher for Sclerotium cepivorum with 71.5 and 62.0%, respectively. The maximum inhibition for Rhizoctonia solani was with fatty hexane extracts of the sapwood (51.4%). Our study suggest that CSC extracts could be used as a possible complementary alternative to synthetic fungicides.

We suggest a deep learning based sensor signal processing method to remove chemical, kinetic and electrical artifacts from ion selective electrodes’ measured values. An ISE is used to investigate the concentration of a specific ion from aqueous solution, by measuring the Nernst potential along the glass membrane. However, application of ISE on a mixture of multiple ion has some problem. First problem is a chemical artifact which is called ion interference effect. Electrically charged particles interact with each other and flows through the glass membrane of different ISEs. Second problem is the kinetic artifact caused by the movement of the liquid. Water molecules collide with the glass membrane causing abnormal peak values of voltage. The last artifact is the interference of ISEs. When multiple ISEs are dipped into same solution, one electrode’s signal emission interference voltage measurement of other electrodes. Therefore, an ISE is recommended to be applied on single-ion solution, without any other sensors applied at the same time. Deep learning approach can remove both 3 artifacts at the same time. The proposed method used 5 layers of artificial neural networks to regress correct signal to remove complex artifacts with one-shot calculation. Its MAPE was less than 1.8% and R² of regression was 0.997. A randomly chosen value of AI-processed data has MAPE less than 5% (p-value 0.016).

Malassezia is lipid-dependent commensal yeast of the human skin. The different culture media and skin sampling methods used to grow these fastidious yeasts are a source of heterogeneity in culture-based epidemiological study results. This study aimed to compare the performances of three methods of skin sampling, and two culture media for the detection of Malassezia yeasts by culture from the human skin. Three skin sampling methods, namely sterile gauze, dry swab and TranswabTM with transport medium, were applied on 10 healthy volunteers. Each sample was further inoculated onto either the novel FastFung medium or the reference Dixon agar for the detection of Malassezia spp. by culture. At least one colony of Malassezia spp. grew on 93/300 (31%) of the cultures, corresponding to 150 samplings. The positive culture rate was 67%, 18%, and 15% (P < 10-3), for samples collected with sterile gauze, TranswabTM, and dry swab, respectively. The positive culture rate was 62% and 38% (P < 0.003) by using the FastFung and the Dixon media, respectively. Our results showed that sterile gauze rubbing skin sampling followed by inoculation on FastFung medium should be implemented in the routine clinical laboratory procedure for Malassezia spp. cultivation.

Unanticipated errors in scientific research data can be attributed to the unwarranted assumption of uniformity in the polystyrene surface that is ubiquitously used in tissue culture flasks and dishes. We have shown that when adherent cells are subjected to fluid shear force, equivalent to rinsing the culture with a balanced salt solution, cells on some areas of the polystyrene surface will immediately rupture while still adherent on the surface. This heterogeneity on the polystyrene surface can cause unexpected variability in experimental results and in replicating experiments among labs. In this paper a novel quantitative method is described to measure the degree of heterogeneity on the polystyrene surface of tissue culture flasks. The results show significant variation among several brands of tissue culture flasks as well as large variability within the production lot of a manufacturer. The assay method involves loading the cells with a fluorescent marker that is released upon membrane rupture. Cell membrane rupture also causes the loss of marker proteins used in Westernblots. This novel assay method can be used to monitor the batch consistency and the manufacturing process of flasks and dishes. It may also be used to test new biomaterials.

Intersectional experiences, socio-cultural meanings, ethnic traditions and morals compound stigma-related stress (Jackson et al., 2020; Schmitz 2019). Sex workers are subject to various stigmatizing forces which can lead to secrecy, isolation and lack of social and cultural support (Koken 2012). Stigmatizing forces include structural humanitarian governance and aid interventions that conflate migration and sex work with insidious constraints and coercion. This study explored how migrant female sex workers from distinctive ethnic cultures manage their identity on a day to day basis in relation to the separation of work and home life. Methods: The perspectives of female sex workers were collected via a series of in-depth semi-structured interviews. The inclusion criteria were that the women had worked in sex work for over 18months, defined their involvement in sex work as voluntary, and were over 18yrs of age. The perspectives of seven women from South Asian (Pakistani), Brazilian, and British backgrounds were analyzed using Grounded Theory (Glaser and Strauss, 1967). Ethnicity was considered to explore how the women experienced stigma, how it impacted on the management of their identity, and how the process of change occurred. Results: The women used a variety of methods to maintain work and home life boundaries, processes they used switch into a role and all experienced stigma and tried to deal with it in ways such as concealment from friends and family. Two core categories and properties emerged from the data as participants felt guilt and/or shame but only the South Asian participants spoke of this with reference to their culture and religion. Conclusion: It was not migration per se but rather the relationship of migration to culture that was key to identity management. Participants reflected that as their country was considered collectivist country with interdependent thought, that any negativity felt could not only be reflected on the individual, but also the entire family. For these reasons Pakistani sex workers were subject to more complex stigmatizing forces, shame and guilt as regards risk and exposure. Discussion focusses on the processes and management strategies used to extend social and cultural support.

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.

Most commercial Cannabis sativa L. (cannabis) genotypes are short-day plants and cultivators typically use a 12.0 h uninterrupted dark period to induce flowering; however, scientific information is lacking to prove this is the optimal dark period for all genotypes, and cultivar specific photoperiods may increase productivity. Tissue culture can be used for research requiring multiple treatments, proper replication, and in a controlled environment on a smaller scale compared to greenhouse and indoor facilities. To determine whether cannabis explants can flower under varied photoperiods in vitro, explants were grown under one of six photoperiod treatments: 12.0, 13.2, 13.8, 14.4, 15.0, and 16.0 h for four weeks. The percentage of flowering explants was highest under 12.0 and 13.2 h treatments. There were no treatment effects on the fresh weight, final height, or growth index of the explants. The results suggest an uninterrupted dark period of at least 10.8 h (i.e. 13.2 h photoperiod) is needed to induce the flowering of this genotype. In vitro flowering could provide a unique and high throughput approach to study floral/seed development and secondary metabolism in cannabis under highly controlled conditions. Further research should determine if this response is the same on a whole plant level.

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.

Many species of the Asteraceae family are used in traditional Mexican medicine for possessing healing properties. Ageratina pichinchensis (Asteraceae) is a plant used for the treatment of gastric ulcers, deep wounds and for its antifungal effects. The aim of this study was to establish a cell suspension culture of A. pichinchensis, quantify the anti-inflammatory constituents 2,3-dihydrobenzofuran and 3-epilupeol, to evaluate the anti-inflammatory potential of its extracts and perform a phytochemical analysis. Cell suspension cultures were established in MS culture medium supplemented with 30 g L-1 sucrose and 1.0 g L-1 α-naphthaleneacetic acid (NAA) plus 0.1 mg L-1 6-furfurylaminopurine (KIN). The ethyl acetate extracts of cell suspension cultures analyzed by GC revealed that the maximum production of compounds The anti-inflammatory activity of these extracts exhibited significant inhibition of NO production. Furthermore, the phytochemical study of EtOAc and MeOH extracts of the biomass on day 20 led to the identification of 17 known compounds. The structures of compounds were assigned by analysis of 1D and 2D NMR data and the remainder by GC-MS. This is the first report of the production of the (-)-Artemesinol, (-)-Artemesinol glucoside, Encecalin and 3,5-diprenyl-acetophenone compounds by a cell suspension cultures of A. pichinchensis.

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.

Hydrogen is useful as a fuel and could be produced by a variety of means. One approach uses artificial photosynthesis where energy from sunlight powers the splitting of water into hydrogen and oxygen. But, biological methods for producing hydrogen has emerged strongly over the past decades. In particular, specific microorganisms could use different substrates to produce hydrogen at differing yields. Such fundamental discoveries with industrial applications thus motivated the use of metabolic engineering approaches and methodologies in enhancing biological hydrogen production through a series of enzyme over-expression, pathway debottlenecking, and gene deletion. However, such approaches heavily rely on the selection of an appropriate microbial chassis for biohydrogen production. With the proper strain in hand, use of alternative substrates may engender greater hydrogen productivities. But learning from the bioprocessing field, co-culture of two compatible microorganisms have been sought after for improving biohydrogen production. In addition, thermophilic microbes may also be useful candidates for exploiting hydrogen production from composting. Future outlook in the field looks into filling our gaps in understanding of the metabolic network that feeds into hydrogen production in different organisms. But, more importantly, problems such as reduced growth rate in engineered microbes point to fundamental issues with using genetically engineered microorganisms for improved biohydrogen production, to which clever bioprocess engineering may yield solutions.

Wolbachia are intracellular endosymbionts of several invertebrate taxa, including insects and nematodes. Although Wolbachia DNA has been detected in ticks, its presence is generally associated with parasitism by insects. To determine whether or not Wolbachia can infect and grow in tick cells, cell lines from three tick species, Ixodes scapularis, Ixodes ricinus and Rhipicephalus microplus, were inoculated with Wolbachia strains wStri and wAlbB isolated from mosquito cell lines. Homogenates prepared from fleas collected from cats in Malaysia were inoculated into an I. scapularis cell line. Bacterial growth and identity were monitored by microscopy and PCR amplification and sequencing of fragments of Wolbachia genes. The wStri strain infected Ixodes spp. cells and was maintained through 29 passages. The wAlbB strain successfully infected Ixodes spp. and R. microplus cells and was maintained through 2-5 passages. A novel strain of Wolbachia belonging to the supergroup F, designated wCfeF, was isolated in I. scapularis cells from a pool of Ctenocephalides sp. cat fleas and maintained in vitro through two passages over nine months. This is the first confirmed isolation of a Wolbachia strain from a flea and the first isolation of any Wolbachia strain outside the “pandemic” A and B supergroups. The study demonstrates that tick cells can host multiple Wolbachia strains, and can be added to panels of insect cell lines to improve success rates in isolation of field strains of Wolbachia.

Family doctors can have an active role in identifying significant population needs and solutions. During COVID-19 epidemic, patient home monitoring with pulse oximetry has been a key aspect of care of patients. However, pandemics bring shortage medical equipment such as pulse oximetry. Through the local maker community in a matter of days four “smart” pulsoximeters were created and built. Following Internet of Things principles, the pulsoximeters were programmed to transmit recorded data through Wi-Fi, in real time, directly to the doctors. Each protype pulsoximeter served a family doctor during the pandemic. Building instructions were shared in maker-oriented websites, potentially leading to additional small-scale productions.

Mammary neoplasm affects a population of intact and elderly female dogs and 50% are malignant. In order to study this disease, cell culture is as a promising preclinical model, creating the opportunity to deposit cell lines at a cell bank, allowing a great reproducibility of the assays and making the validation of the results more reliable. Another important aspect is the possibility to establish models for better understanding tumour characteristics, such as vasculogenic mimicry. Due the importance of cancer cell lines in preclinical models, this study aimed to establish and characterize primary cell lines from canine mammary gland tumours according to immunophenotype and tumorigenicity, and with its ability to form vasculogenic mimicry-like structures in vitro and in vivo. Cell cultures were evaluated for morphology, phenotype, vasculogenic mimicry and tumorigenicity abilities. We collected 17 primary mammary carcinoma and 3 metastasis and had a satisfactory result in 10 of them. All cell lines presented spindle shape or polygonal morphology and expressed concomitant pan-cytokeratin and cytokeratin 8/18. Four cell lines had vasculogenic mimicry ability in vitro and two of them showed in vivo tumorigenic potential and forming VM in the xenotransplant tumour. Cell characterization of those lines will help to create a database for more knowledge of mammary carcinomas in dogs, including studies of tumour behaviour and new therapeutic targets.

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.

Purpose: "Just culture" is an element of safety culture, and in a broader sense – a part of quality culture. It is the subject of studies, especially in healthcare. This phenomenon is almost unknown in Polish medical facilities. For this reason, the aim of the article is to present the essence and significance of "just culture" in healthcare. The other aim of the research is to present the results of the validation of "just culture" assessment instrument used to recognize the "just culture" maturity level and evaluate the nurses’ beliefs and behaviours in the light of "just culture" criteria. Methodology/Approach: The verified questionnaire consisted of 28 statements in relation to which respondents expressed their opinion on a 5-point Likert scale. The questionnaire was distributed among nurses in one of the largest hospitals in Pomorskie Voivodeship, in Poland. The results based on 68 responses were statistically processed with Statistica 13.1 software. Findings: The obtained results allowed to confirm the reliability of the assessment tool, to recognize the level of „just culture” as wisdom (68%) and to indicate strengths and weaknesses of observed beliefs and behaviours. On this basis, improvement actions were proposed. Originality/Value: We use the original, own prepared questionnaire. This is the first study on "just culture" in healthcare in Poland. There are only few studies devoted to patient safety culture in Poland and no research addressed to "just culture" phenomenon, as well in Poland as in Central Europe. The results in this area allow to recommend the assessment tool for other hospitals and seem to help in understanding the essence of "just culture" implementation.

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.

Contact-based co-culture of fibroblasts and keratinocytes is important to study the structure and functions of the wound bed. Co-culture of these two cell types in direct contact with each other has been challenging, requiring high serum concentrations (up to 10%), feeder systems and a range of supplemental factors. These approaches are not only technically demanding, but also present scientific, cost and ethical limitations associated with high-serum concentrations. We have developed two reduced-serum approaches (1-2%) to support contact-based co-culture of human dermal fibroblasts (HDFa) and human epidermal keratinocytes (HaCaT). The two approaches include (1) Specialized cell culture media for each cell type mixed in a 1:1 ratio (KGM+FGM), and (2) Minimal media supplemented with cell-specific growth factors (MEM+GF). Co-culture could be successfully achieved by co-seeding (two cell types were introduced simultaneously), or in a layered fashion (keratinocytes seeded on top of confluent fibroblasts). With wound scratch assays, the co-cultured platforms could demonstrate cell proliferation, migration and wound closure. The reduced-serum conditions developed are simple, easy to formulate and adopt, and based on commonly-available media components. These contact-based co-culture approaches can be leveraged for wound and skin studies, and tissue bioengineering applications, potentially reducing concerns with high-serum formulations.

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.

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.

Biofilm-mediated infection is a major cause of bone prosthesis failure. The lack of molecules able to act in biofilms has driven research aimed at identifying new anti-biofilm agents via chemical screens. However, to be able to accommodate a large number of compounds, the testing conditions of these screenings end up being typically far from the clinical scenario. In this study, we assess the potential applicability of three anti-biofilm compounds (based on natural compounds) as part of implanted medical devices by testing them on in vitro systems that more faithfully resemble the clinical scenario. To that end, we used a competition model based on the co-culture of SaOS-2 mammalian cells and Staphylococcus aureus (collection and clinical strains) on a titanium surface. Additionally, we studied whether these derivatives of natural compounds enhance the previously proven protective effect of pre-incubating the titanium surface with SaOS-2 cells. Out of the three tested leads, one showed the highest potential, and can be regarded as a promising agent for incorporation into bone implants. This study emphasizes and demonstrates the importance of using meaningful experimental models, where potential antimicrobials ought to be tested for protection of biomaterials in translational applications.

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.

Increasing pesticide resistance in plant pathogens is major concern in agriculture production. Research on ecofriendly alternatives of chemical pesticides are more in demand in pesticide industry. In the current study, an ethyl acetate extract from the endophytic fungus Trichoderma sp. EFI 671, isolated from the stem parts of the medicinal plant Laurus sp., was screened for bioactivity against plant pathogens (Fusarium graminearum, Rhizoctonia solani, Sclerotinia sclerotiorum and Botrytis cinerea), insect pests (Spodoptera littoralis, Myzus persicae, Rhopalosiphum padi) and plant parasites (Meloidogyne javanica). The bioactive components have been characterized following a bioassay-guided isolation against M. persicae. The chemical study of this bioactive extract resulted in the isolation of 1-oleoyl-2-linoleoyl-3-palmitoylglycerol (1), eburicol (2), (24R)-stigmast-4-ene-3-one or β-sitostenone (3), ergosterol (4) and ergosterol peroxide (5). The free fatty acids present in compound 1 (oleic, linoleic and palmitic) showed strong dose-dependent aphid antifeedant effects against M. persicae. Liquid (PDB, and SDB) and solid (corn, sorghum, pearl millet and rice) growth media were tested in order to optimize the yield and bioactivity of the fungal extracts. Pearl millet and corn gave the highest extract yields. All the extracts from these solid media had strong effects against M. persicae with sorghum being the most active. Corn increased the content in linolenic, pearl millet the oleic and stearic and sorghum oleic and linolenic acids compared to rice. Their antifeedant effects correlated with linoleic /oleic acids. The phytotoxic effects of these extracts against Lolium perenne and Lactuca sativa varied with culture media, with sorghum being the least toxic.

The purpose of this work is to define optimal growth conditions for batch culture of the cyanobacterium Arthrospira maxima and the microalgae Chlorella vulgaris, Isochrysis galbana and Nannochloropsis gaditana. Thus, we study the effect of three variables on algae growth: i.e., inoculum:culture medium ratio, light:darkness photoperiod and type of culture medium, including both synthetic media and wastewaters. The results showed that the initial inoculum volume did not affect the amount of biomass at the end of the growth (14 days), whereas an excess (18 h) or defect (6 h) in the number of hours of light is determinant for its development. The contribution of nutrients from different culture media modified the growth of the different species. A. maxima was favoured in seawater enriched with Guillard's F/2 as well as C. vulgaris and N. gaditana but in fresh water medium. I. galbana had the greatest growth in the marine environment enriched with Walne’s media. Nitrate was the limiting growth reagent at the end of the exponential phase of growth for C. vulgaris and N. gaditana, while iron was for A. maxima and I. galbana. All species demonstrated their capability to grow in effluents from a wastewater treatment plant and they efficiently consume nitrogen, especially the three microalgae species.

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.