Background. Currently, most of the research in breast cancer has been carried out in conventional two-dimensional (2D) cell culture due to its practical benefits, however, the three-dimensional (3D) cell culture is becoming the model of choice in cancer research because it allows cell-cell and cell-extracellular matrix (ECM) interactions, mimicking the native microenvironment of tumors in vivo. Methods. in this work, we evaluated the effect of 3D cell organization on the expression pattern of miRNAs (by Small-RNAseq) and mRNAs (by microarrays) in the breast cancer SKBR3 cell line and analyzed the biological processes and signaling pathways regulated by the differ-entially expressed protein-coding genes (DE-mRNAs) and miRNAs (DE-microRNAs) found in the organoids. Results. We obtained well-defined cell-aggregated organoids with a grape cluster-like morphology with a size up to 9.2×105 μm3. The transcriptomic assays showed that cell growth in organoids significantly affected (all p < 0.01) the gene expression patterns of both, miRNAs and mRNAs, finding 20 upregulated and 19 downregulated DE-microRNAs, as well as 49 upregulated and 123 downregulated DE-mRNAs. In silico analysis showed that a subset of 11 upregulated DE-microRNAs target 70 downregulated DE-mRNAs. These genes are involved in 150 gene on-tology (GO) biological processes such as regulation of cell morphogenesis, regulation of cell shape, regulation of canonical Wnt signaling pathway, morphogenesis of epithelium, regulation of cy-toskeleton organization, as well as in the MAPK and AGE-RAGE signaling KEGG-pathways. In-terestingly, hsa-mir-122-5p (Fold Change (FC)=15.4), hsa-mir-369-3p (FC=11.4), and hsa-mir-10b-5p (FC=20.1) regulated up to the 81 % of the 70 downregulated DE-mRNAs. Conclu-sion, the organotypic 3D cell-organization architecture of breast cancer SKBR3 cells impacts the expression pattern of miRNAs-mRNAs network mainly through overexpression of hsa-mir-122-5p, hsa-mir-369-3p, and hsa-mir-10b-5p. All these findings suggest that the interaction between cell-cell and cell-ECM as well as the change in the culture architecture impacts gene ex-pression, and therefore, support the pertinence of migrating breast cancer research from conven-tional cultures to 3D models.

The process of identifying and approving a new drug is a time-consuming and expensive procedure. One of the biggest issues to overcome is the risk of hepatotoxicity, which is one of the main reasons for drug withdrawal from the market. While animal models are the gold standard in preclinical drug testing, the translation of results into therapeutic intervention is often ambiguous due to interspecies differences in hepatic metabolism. The discovery of human induced Pluripotent Stem Cells (hiPSCs) and their derivatives has opened new possibilities for drug testing. We used mesenchymal stem cells and hepatocytes both derived from hiPSC, together with endothelial cells, to miniaturize the process of generating hepatic organoids. These organoids were then cultivated in vitro using both static and dynamic cultures. Additionally, we tested spheroids composed solely by induced hepatocytes. By miniaturizing the system, we demonstrated the possibility of maintaining the organoids, but not the spheroids, in culture for up to 15 days. This timeframe may be sufficient to carry out a hypothetical pharmacological test or screening. In conclusion, we propose that the hiPSC-derived liver organoids model could complement or, in the near future, replace the pharmacological and toxicological tests conducted on animals.

Malignant cells cultured in three-dimensional (3D) models have been found to be phenotypically and biochemically different from their counterparts cultured conventionally. Since most of these studies employed solid tumor types, how 3D culture affects multiple myeloma (MM) cells is not well understood. Here, we compared MM cells (U266 and RPMI8226) in a 3D culture model with those in conventional culture. While the conventionally cultured cells were present in single cells or small clusters, MM-3D cells grew in large spheroids. We discovered that STAT3 was the pathway that was more activated in 3D in both cell lines. The active form of STAT3 (phospho-STAT3 or pSTAT3), which was absent in MM cells cultured conventionally, became detectable after 1-2 days in 3D culture. This elevated pSTAT3 level was dependent on the 3D environment, since it disappeared after transferring to conventional culture. STAT3 inhibition using a pharmacological agent, Stattic, significantly decreased the cell viability of MM cells and sensitized them to bortezomib in 3D culture. Using an oligonucleotide array, we found that 3D culture significantly increased the expression of several known STAT3 downstream genes implicated in oncogenesis. Since most primary MM tumors are naturally STAT3-active, studies of MM in 3D culture can generate results that are more representative of the disease.

We previously designed and synthesized dehydroxyepoxyquinomicin (DHMEQ) as an inhibitor of NF-κB based on the structure of microbial secondary metabolite epoxyquinomicin C. DHMEQ showed anti-inflammatory and anticancer activity in various in vivo disease models without toxicity. Cell detachment from the primary tumor and subsequent invasion are considered to be early phase of metastasis, while tumor cell attachment to the tissue and secondary tumor formation the late phase. The assay system for late phase was set up with intra-portal-vein injection of pancreatic cancer cells. Administration of DHMEQ was found to inhibit the liver metastasis possibly by decreasing the expression of MMP-9 and IL-8. Also when the pancreatic cancer cells treated with DHMEQ was inoculated into the peritoneal cavity of mice, the metastatic foci formation was inhibited. These results indicate that DHMEQ is likely to inhibit the late phase of metastasis. Meanwhile, we have recently employed three-dimensional (3D) culture of breast cancer cells for the model of early phase metastasis. DHMEQ inhibited the 3D invasion of breast cancer cells without toxicity. In this way, DHMEQ was shown to inhibit the late and early phases of metastasis. Thus, DHMEQ is likely to be useful for the suppression of cancer metastasis.

We are witnessing the revival of the CAM model, which was already used in the past by some authors studying angiogenesis and anti-cancer drugs, that now offers a refined model to fill, in the translational meaning, the gap between in vitro-in vivo studies. It can be used for a wide range of purposes, from testing cytotoxicity, pharmacokinetics, tumorigenesis, invasion, to action mechanisms of molecules, and validation of new materials from tissue engineering research. CAM model is easy to use with a fast outcome and makes experimental research more sustainable since allowing to replace, reduce and refine pre-clinical experimentation ("3Rs" rules [1]). This re-view aims to highlight some unique potentials that CAM-assay presents; in particular, the authors intend to use in the next future the CAM model to verify, in a microenvironment comparable to in vivo conditions, albeit simplified, the angiogenic ability of functionalized 3D constructs to be used in regenerative medicine strategies in the recovery of skeletal injuries of critical size (CSD) that do not repair spontaneously. For this purpose, organotypic cultures will be planned on sever-al CAMs set up in temporal sequence: a sort of organ model for assessing CSD conducted in the CAM bioreactor, rather than in vivo.

Using a combination of pan proteomic platform associated with systemic biology analyses, we demonstrate that neonatal microglial cells derived from cortex and spinal cord expressed different phenotypes upon the physiological or pathological conditions. They also highlight great variability in protein production on both cellular and exosome levels. Bioinformatics data indicate for the cortical microglia anti-inflammatory and neurogenesis/tumorigenesis characteristics, while for the spinal cord microglia involvement in the inflammatory response. We confirmed these results by performing functional testing including neurite outgrowth assays in DRGs cell line, and glioma proliferation analysis in 3D spheroid cultures. Results from these in vitro assays indicate that the microglia located at different CNS areas reveal differential biological functions. While both microglia sources enhanced growth of DRGs axons, only the spinal microglia significantly attenuated glioma proliferation. Overall these findings are pointing to the fact that the origin of neonatal microglia affects the physio-pathological function, which may address the prevalence of the glioma in the brain in comparison with the spinal cord in adult.

Prior to clinical trials, preclinical testing of oncology drug candidates is performed by evaluating drug candidates with in vitro and in vivo platforms. For in vivo testing, animal models are used to evaluate the toxicity and efficacy of drug candidates. However, animal models often display poor translational results as many drugs that pass preclinical testing fail when tested in humans, with oncology drugs exhibiting especially poor acceptance rates. The FDA Modernization Act 2.0 promotes alternative preclinical testing techniques, presenting the opportunity to use higher complexity in vitro models as an alternative to in vivo testing, including 3D cell culture models. 3D tissue cultures address many of the shortcomings of 2D cultures by more closely replicating the tumour microenvironment through a combination of realistic drug diffusion, paracrine signaling, cellular phenotype, and vascularization that can better mimic native human tissue. This review will discuss the common forms of 3D cell culture, including cell spheroids, organoids, organs-on-a-chip, and 3D bioprinted tissues. Their advantages and limitations will be presented, aiming to discuss the use of these 3D models to accurately represent human tissue and as an alternative to animal testing. The use of 3D culture platforms for preclinical drug development is expected to accelerate as these platforms continue to improve in complexity, reliability, and translational predictivity.

Culture systems for 3-dimensional tissues, such as multicellular spheroids, are indispensable for high-throughput screening of primary or patient-derived xenograft (PDX)-expanded cancer tissues. Oxygen supply to the center of such spheroids is particularly critical for maintaining cellular functions as well as avoiding the development of a necrotic core. In this study, we evaluated 2 methods to enhance oxygen supply: (1) using culture plate with gas-permeable polydimethylsiloxane (PDMS) membrane at its bottom and (2) embedding hydrogel beads in the spheroids. Culturing spheroids on PDMS increased cell growth and affected glucose/lactate metabolism and CYP3A4 mRNA expression and subsequent enzyme activity. The spheroids comprised 5000 Hep G2 cells and 5000 20 µm-diameter hydrogel beads did not develop a necrotic core for 9 days when cultured on a gas-permeable sheet. In contrast, central necrosis in spheroids lacking hydrogel beads was observed after day 3 of culture, even when using PDMS. These results indicate that the combination of gas-permeable culture equipment and embedded hydrogel beads improves culture 3D spheroids produced from primary or PDX-expanded tumor cells.

Glioblastoma (GBM), the most common and aggressive primary brain tumor in adults, remains one of the least treatable cancers. Current standard of care—combining surgical resection, radiation, and alkylating chemotherapy—results in a median survival of only 15 months. Despite decades of investment and research into the development of new therapies, most candidate anti-glioma compounds fail to translate into effective treatments in clinical trials. One key issue underlying this failure of therapies that work in pre-clinical models to generate meaningful improvement in human patients is the profound mismatch between drug discovery systems—cell cultures and mouse models—and the actual tumors they are supposed to imitate. Indeed, current strategies that evaluate the effects of novel treatments on GBM cells in vitro fail to account for a wide range of factors known to influence tumor growth. These include secreted factors, the brain’s unique extracellular matrix, circulatory structures, the presence of non-tumor brain cells, and nutrient sources available for tumor metabolism. While mouse models provide a more realistic testing ground for potential therapies, they still fail to account for the full complexity of tumor-microenvironment interactions, as well as the role of the immune system. Based on the limitations of current models, researchers have begun to develop and implement novel culture systems that better recapitulate the complex reality of brain tumors growing in situ. A rise in the use of patient derived cells, creative combinations of added growth factors and supplements, may provide a more effective proving ground for the development of novel therapies. This review will summarize and analyze these exciting developments in 3D culturing systems. Special attention will be paid to how they enhance the design and identification of compounds that increase the efficacy of radiotherapy, a bedrock of GBM treatment.

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.

Mycotoxins are secondary metabolites produced by filamentous fungi associated to a variety of acute and chronic foodborne diseases. Current toxicology studies mainly rely on monolayer cell cultures and animal models, which are undeniably affected by several limitations. To bridge the gap between the current in vitro toxicology approach and the in vivo predictivity of the data, we here investigated the cytotoxic effects induced by the mycotoxins sterigmatocystin (STE), ochratoxin A (OTA) and patulin (PAT) on different 2D and 3D cell cultures. We focused on human tumours (neuroblastoma SH-SY5Y cells, and epithelial breast cancer MDA-MB-213 cells) and healthy cells (bone marrow derived mesenchymal stem cells, BM-MSC, and umbilical vein endothelial cells, HUVECs). Cytotoxicity of STE, OTA, and PAT was determined after 24, 48 and 72 h of exposure by ATP assay in both culture models. 3D spheroids morphology was also analysed using the MATLAB-based open-source software AnaSP. Our results highlighted how each cell line and different culture models showed specific sensitivities, reinforcing the importance of using more complex models for toxicology studies and a multiple cell line approach for an improved and more comprehensive risk assessment.

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.

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.

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

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

The 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.

Based on a literature review, a framework for digital organizational culture with a focus on innovation is proposed. This digital organizational culture becomes relevant today since digital transformation requires an appetite for risk and a willingness to undertake, aspects that are strengthened by an environment and atmosphere that motivate individuals to generate new ideas. The proposal begins with a diagnosis of the existing organizational culture, which allows identifying the differences between the components of the current organizational culture and the innovation-oriented digital organizational culture. Through this diagnosis, the gaps that need to be addressed to achieve a digital organizational culture with innovative characteristics are identified. In addition, specific recommendations are proposed to carry out this transformation. In the process, the assumptions underlying the current typology of the company's organizational culture are highlighted. The importance of integrating creativity, risk readiness, and entrepreneurial willingness into the digital organizational culture aimed at innovation is emphasized. These elements are essential to create an environment

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.

Three-dimensional (3D) cell culture systems can be regarded as suitable platforms to bridge the huge gap between animal studies and two-dimensional (2D) monolayer cell culture to study chronic diseases such as cancer. In particular, the preclinical platforms for multicellular spheroid formation and culture can be regarded as ideal in vitro tumor models. The complex tumor microenvironment such as hypoxic region and necrotic core can be recapitulated in 3D spheroid configuration. Cells aggregated in spheroid structures can better illustrate the performance of anti-cancer drugs as well. Various methods have been proposed so far to create such 3D spheroid aggregations. Both conventional techniques and microfluidic methods can be used for generation of multicellular spheroids. In this review paper, we first discuss various spheroid formation phases. Then, the conventional spheroid formation techniques such as bioreactor flasks, liquid overlay and hanging droplet technique are explained. Next, a particular topic of the hydrogel in spheroid formation and culture is explored. This topic has received less attention in the literature. Hydrogels entail some advantages to the spheroid formation and culture such as size uniformity, the formation of porous spheroids or hetero-spheroids as well as chemosensitivity and invasion assays and protecting from shear stress. Finally, microfluidic methods for spheroid formation and culture are briefly reviewed.

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.

Carbapenemase resistance in Enterobacterales is a global public health problem and rapid and effective methods to detect resistance mechanisms are needed urgently. Our aim was to evaluate the performance of a MALDI-TOF MS based KPC detection protocol from patients’ positive blood cultures, short-term cultures and colonies at health care settings. Bacterial identification and KPC detection were achieved after protein extraction with organic solvents and target spot loading with suitable organic matrices. Confirmation of KPC production was performed by susceptibility tests, blaKPC amplification by PCR and sequencing. KPC direct detection (KPC-peak at approximately 28.681 Da) from patients’ positive blood cultures, short-term cultures and colonies, once bacterial identification was achieved, showed an overall sensibility and specificity of 100% (CI95: [95%,100%] and CI95: [99%, 100%], respectively). Concordance between hospital routine bacterial identification protocol and identification with this new methodology from the same extract used for KPC detection was ≥92%. This study represents the pioneering effort to directly detect KPC using MALD-TOF MS technology, conducted on patient-derived samples obtained at the hospitals for validation purposes, in a multi-resistance global context that requires concrete actions to preserve available therapeutic options and reduce the spread of antibiotic resistance markers.

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.

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

In the bioprinting process, controlling the motion of bioprinters involves a computer-aided design (CAD) model, converting that model into g-code, and transmitting the motion commands to the bioprinters. The g-code file contains information about the motion of the axes and can be generated using various software. However, a challenge arises when using g-code between different bioprinters. Therefore, the objective of this study was to propose a reliable method for analyzing the differences in 3D bioprinting devices by parameterizing a crucial variable for extrusion bioprinting: the E-value. Four bioprinters from three different manufacturers were used to investigate the extrusion behavior when subjected to the same E-value.

Cell-culture meat is a novel concept to replace traditional meat produced from animals, and to satisfy rising food demands due to the world population growth. It is still a young field and requires to solve technical issues including cell sources and types, efficient culture for large scale production, taste and texture for customer satisfaction. The current review summarized traditional beef meat production and evaluation methods for customer satisfaction. In particular, serum biomarkers for beef meat evaluation are discussed, which can be applicated for cell-culture meat production.

Taking into account the growing interest in microalgae to be used as raw material for biodiesel production, this research is aimed at analyzing the rheological behaviour of microalgae suspensions (Chlorella sp) at different culture times under eight different conditions (temperature, salinity and CO₂, NO₃ and PO₄ levels) in order to estimate the energy demands of each step, with the purpose of optimizing a continuous feed tubular bioreactor construction. For each condition, it was calculated the biomass and oil yields, so as to correlate these results with rheological parameters. The suspension results indicated that the microalgae Chlorella sp is a non-Newtonian material with dilatant characteristics; the processing time hardly exerted an influence on the rheograms of the suspension of the microalgae Chlorella sp, except for the simultaneous conditions of low salinity and low CO₂ content; NO₃ and PO₄ contents and the amount of supplements influenced the rheological parameters of the suspension of the microalgae Chlorella sp, when in low concentration of CO₂ and low salinity levels.

Three-dimensional (3D) printing technology, also known as additive manufacturing (AM), has emerged as an attractive state-of-the-art tool for precisely fabricating functional materials with complex geometries, championing several advancements in tissue engineering, regenerative medicine, and therapeutics. However, this technology has an untapped potential for biotechnological applications, such as sensor and biosensor development. By exploring these avenues, the scope of 3D printing technology can be expanded and pave the way for groundbreaking innovations in the biotechnology field. Indeed, new printing materials and printers would offer new possibilities for seamlessly incorporating biological functionalities within the growing 3D scaffolds. Herein, we review the extrusion-based 3D printing modalities emphasizing biosensor applications. We highlighted the application of natural, synthetic, and composite biomaterials as 3D-printed soft hydrogels. Emphasis was placed on the approach by which the sensing molecules were introduced during the fabrication process. Finally, the future perspective was provided.

Synthesis of heterodinuclear 3d transition metal complexes in which two metal ions are introduced in equivalent coordination sites of a ligand is quite uncommon. We have previously shown that the application of the cis-trans isomerization of the oxamido group was able to furnish such a possibility. In the present paper we demonstrate that this synthetic pathway, relying on the possibility of changing the conformation (cis vs trans) of the oxamido group, can be extended to other ligands. Although we have not been able to structurally characterize these complexes, EPR and magnetic data do confirm the efficiency of this reaction pathway. We also demonstrate that the cis-trans isomerization of the oxamido group is not due to a particular property of the ion involved in the reaction. In fact, isomerization is governed by the basicity of the reaction medium. According to the reaction condition, it is possible to favor or not this phenomenon, and to increase the accessibility to a larger number of novel genuine complexes. But the oxophilic character of the lanthanide ions impedes this isomerization in 3d-4f complexes. The very weak magnetic interactions observed in these Cu-Gd complexes can be explained thanks to spin polarization.

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.

Based on three-dimensional optical proximity correction (3D OPC), recent advancements in 3D lithography have enabled the high-fidelity customization of 3D micro-optical elements. However, micron-to-millimeter scale structures represented by Fresnel lens design bring more stringent requirements for 3D OPC, which poses significant challenges to the accuracy of models and the efficiency of algorithms. Thus, a lithographic model based on optical imaging and photo-chemical reaction curve is developed in this paper, and a sub-domain division method with a statistics principle is proposed to improve the efficiency and accuracy of 3D OPC. Both the simulation and the experimental results showed the superiority of the proposed 3D OPC method in the fabrication of the Fresnel lenses. The computation memory requirements of the 3D OPC are reduced to below 1 %, and the profile error of the fabricated Fresnel lens reduced 79.98%. Applying the Fresnel lenses to an imaging system, the average peak signal to noise ratio (PSNR) of the image increased by 18.92 %, and the average contrast of the image enhanced by 36 %. We believe the proposed 3D OPC method can be extended to the fabrication of vision-correcting ophthalmological lenses.

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.

While Neural Radiance Fields (NeRF) are gaining increasing interest in various domains as innovative methods for novel view synthesis and image-based reconstruction, their potential application in the realm of Cultural Heritage remains unexplored. Purpose of this paper is to assess the effectiveness of applying NeRF to sets of images of digital heritage objects and sites. The study’s findings demonstrate that NeRF could be valuable when used in combination with or as a comparison to other well-established techniques such as photogrammetry, to expand the possibilities of preserving and presenting heritage assets with enhanced visual fidelity and accuracy. Particularly, NeRF show promising results in improving the rendering of translucent and reflective surfaces, objects with homogeneous textures, and elements with intricate details. In addition, we demonstrate that, when considering the same set of input images (with known camera poses), reducing the image quality or the number of images results in significantly less information loss with NeRF compared to photogrammetry. This suggests that NeRF is preferentially suited for scenarios involving sparse information or low-quality photos or videos, which could be especially valuable in risky or challenging situations.

3D printing is playing an important role in cardiovascular disease by use of personalised models with replication of normal anatomy and pathology with high accuracy and reliability. While 3D printed heart and vascular models are shown to improve medical education, preoperative planning and simulation of cardiac procedures, and enhance communication with patients, 3D bioprinting however, represents a potential advancement of 3D printing technology by printing cellular or biological components, functional tissues and organs that can be used in a variety of applications in cardiovascular disease. Recent advances in bioprinting technology have shown the ability to support vascularisation of large scale constructs with enhanced biocompatibility and structural stability, thus creating opportunities to replace damaged tissues or organs. In this review, we provide an overview of the use of 3D bioprinting in cardiovascular disease with a focus on technologies and applications in cardiac tissues, vascular constructs and grafts, heart valves and myocardium. Limitations and future research directions are highlighted.

Tumor cells evolve in a complex and heterogeneous environment composed of different cell types and an extracellular matrix. Current 2D culture methods are very limited in their ability to mimic the cancer cell environment. In recent years, various 3D models of cancer cells have been developed, notably in the form of spheroids/organoids, using scaffold or cancer-on-chip devices. However, these models have the disadvantage of not being able to precisely control the organization of multiple cell types in complex architecture and are sometimes not very reproducible in their production, and this is especially true for spheroids. Three-dimensional bioprinting can produce complex, multi-cellular, and reproducible constructs in which the matrix composition and rigidity can be adapted locally or globally to the tumor model studied. For these reasons, 3D bioprinting seems to be the technique of choice to mimic the tumor microenvironment in vivo as closely as possible. In this review, we discuss different 3D-bioprinting technologies, including bioinks and crosslinkers that can be used for in vitro cancer models, and the techniques used to study cells grown in hydrogels; finally, we provide some applications of bioprinted cancer models.

Background: The main objective of this study is to develop a customized metatarsophalangeal implant for the treatment of various conditions affecting this specific joint. This approach involves replacing the joint with a custom implant designed specifically to fit each patient's unique anatomical structure. The need for custom implants stems from the obvious lack of size variety available, which presents a major challenge when, the search for the ideal size implant can be a hard task, also when considering the possibility of extensive modification of the bone to adapt it to the available implant. This adaptation, often synonymous with bone reduction, can cause significant post-operative pain for the patient and impact healing time. Methods: A comparative study of modeling software was carried out to select the perfect software, by defining and comparing the relevant criteria. Similarly, the different biomaterial options were examined and compared to select the one best suited to this project, mentioning the imaging modality used to determine the types of images used for our approach. Finally, the 3D printers used in this work were represented, highlighting their technical characteristics. Findings: A resin metatarsophalangeal implant prototype was created using 3D modeling and reconstruction software and tools. Interpretation: Reconstruction and 3D modeling techniques are revolutionizing implant creation, offering customized solutions for patients suffering from metatarsophalangeal joint disorders caused by a wide range of diseases. These results have significant implications for disability management and rehabilitation, encouraging further exploration and progress.

The endorsement from Christian circles in facilitating the rise of right-wing populism in Brazil, the United States, as well as the support and alliance of the Polish Catholic Church with the Law and Justice (PiS) government, is widely acknowledged. Embedded within the intricate fabric of multifaceted factors contributing to the surge of right-wing populism, the Culture War assumes a pivotal yet often underestimated role. Manifesting as a profound confrontation, the Culture War signifies an existential clash between contrasting perspectives concerning socio-cultural values grounded in religious scripture. Progressive endeavors strive to deconstruct longstanding norms, advocating for the ascendancy of post-materialistic ideals, while conservative domains portray an unwavering commitment to safeguarding national and religious socio-cultural values as sacrosanct. Both factions employ moral-ethical justifications in their respective pursuits, rendering the landscape of cultural discourse a nuanced tapestry of ideological contention. The influence of Christian doctrine on the conflict surrounding social justice, and the potential convergence between Christian doctrine and the positions of Populist right-wing movements on social justice issues, is underscored. By employing a singular case study focused on Portugal, owing to the nation's enduring relationship with Catholicism spanning from Monarchical times to the Republic and the Fascist Catholic regime, Portugal emerges as indisputably one of the most religious countries in the Western world, akin to Poland. Consequently, in theory, the support of the Portuguese Catholic Church for the nascent Populist right-wing and Christian-oriented party should be evident amidst the Culture War conflict, wherein conservative religious Christian values face existential challenge. However, the backing extended to CHEGA by both the Catholic Church and the Portuguese Episcopal Conference assumes an ambivalent and markedly diminished stance, diverging from the previously robust support observed in established center-right mainstream parties such as the PSD and CDS. Despite this reality and the absence of official support for CHEGA, numerous Christian politicians or activists have expressed endorsement or alignment with the right-wing party, departing from their former affiliations, primarily due to social justice concerns.

Microalgae have emerged as potential feed stock for third generation biofuel and other bioactive compounds. In the present investigation we have evaluated the two native strains of Oscillatoria (O. subbrevis MTC-20702 and O. sancta MTC-20703) isolated from two different fresh water biotopes of Meerut, U.P., India and were subjected for the characterization of generation time, biomass, bio-molecules including Chlorophyll-a, carotenoids, carbohydrate, lipids, phycobilins and proteins. Both strains of Oscillatoria were grown in batch culture till the initiation of declining phase and were harvested on every 10th day of incubation till the starting of declining phase. Result of the present investigation revealed that O. subbrevis MTC-20702 can be potentially exploited for the production of high algal biomass, feed stock for biofuel, Chl-a, carotenoids, c-Allophycocyanin and maximum yield of these bioactive compounds can be achieved on 20th day. However, O. santa could be very good source for c-Phyocyanin, c-Phycoerythrin, carbohydrate and protein and these also can be achieved highest on 20th day of incubation. In addition, as these two strains were isolated from water reservoirs and they can be cultivated in large scale for the production of desired bio-molecules.

Artificial Intelligence is developing at a rapid place, with examples of Machine Learning already utilized in aviation to improve efficiency. In the coming decade it is likely that Intelligent Assistants (IAs) or ‘Digital Colleagues’ will be developed and deployed to assist aviation personnel in the cockpit, the air traffic control centre and airports. This will be a game-changer and may herald the way forward for single pilot operations, AI-based management of Urban Mobility including fleets of drones and sky-taxis, and more carbon-efficient transport across global commercial transport networks. Yet in aviation there is a core underlying tenet that ‘people make safety’ and keep the skies and passengers safe, based on a robust industry-wide safety culture. The introduction of IAs into aviation could potentially undermine aviation’s hard-won track record in this area. This paper applies a scientifically validated safety culture tool to explore the potential impacts of the introduction of IAs into aviation. The results suggest that there are indeed potential negative outcomes, but also possible safety ‘affordances’ wherein AI could strengthen safety culture. Safeguards and mitigations are suggested for the key ‘risk owners’ in aviation organisations, to ensure safety remains a priority in the industry.

. Gut microbiome reflects healthy and predicts possible disease in hosts. A holistic view of this community is needed, focused on identification species, and on the dissection of how species interact with their host and with each other, regardless of whether their presence is beneficial, inconsequential, or detrimental. Distribution of gut-associated eukaryotes within and across NHPs is likely driven by host behavior and ecology. To ascertain the existence of free-living amoebae (FLA) in the gut of wild and captive non-human primates (NHPs), 01 sample stools were collected and submitted to culture-dependent and microscopy examination and DNA sequencing. FLAs were detected in 45.4% (46/101) of fecal samples analysed and their morphological characteristics matching those of Acanthamoeba spp, Vermamoeba spp, and fan-shaped amoebae of the family Vannellidae. Sequence analysis of the PCR- products revealed that the suspected amoebae are highly homologous (99% identity and 100% query coverage) with Acanthamoeba genotype T4 and Vermamoeba vermiformis amoebae. The results showed a great diversity of FLAs in the NHP microbiome, which may pose a potential risk to the health of NHPs. To the best of our knowledge, this is the first report the presence of FLAs in NHPs naturally infected. However, it is not known whether gut-borne FLA are exploiting a viable ecological niche, or whether they are simply transient residents in the gut.

Propagation of human cytomegalovirus (CMV) in cultured cells results in genetic adaptations that confer improved growth in vitro and significant attenuation in vivo. Mutations in RL13 arise quickly during cell culture passage, while mutations in the UL128-131A locus emerge later during fibroblast passage and disrupt expression of a glycoprotein complex that is important for entry into epithelial and endothelial cells. As in vivo CMV replicates in the context of host antibodies, we reasoned that antibodies might mitigate the accumulation of adaptive mutations during cell culture passage. To test this, CMV in infant urine was used to infect replicate fibroblast cultures. One lineage was passaged in the absence of CMV-hyperimmuneglobulin (HIG) while the other was passaged with HIG in the culture medium. The former lost epithelial tropism and aquired mutations disrupting RL13 and UL131A expression, whereas the latter retained epithelial tropism and both gene loci remained intact after 22 passages. An epitheliotropic RL13+/ UL131A+ virus was isolated by limiting-dilution in the presence of HIG and expanded to produce a working stock sufficient to conduct cell tropism experiments. Thus, culture in the presence of antibodies may facilitate in vitro experiments using viruses that are genetically more authentic than has been previously possible.

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

The present study aimed to characterize the microstructure of a temporary 3D printing resin (Resilab Temp), evaluating the optical properties and mechanical behavior according to different post-processing curing times. For the analysis of the surface microstructure and establishment of the best printing pattern, samples in bar format following ISO 4049 (25 x 10 x 3 mm) were designed in CAD software (Rhinoceros 6.0), printed on the W3D printer (Wilcos), and light-cured in Anycubic Photon at different times (without curing, 16 minutes, 32 minutes, and 1 hour). For the structural characterization, analyses were carried out in FTIR (Fourier Transform Infrared Spectroscopy) and Scanning Electron Microscopy (SEM). The mechanical behavior of the resin was based on flexural strength tests and Knoop microhardness. Color and translucency analysis were performed with a spectrophotometer (VITA Easy Shade Advanced 4.0), which was evaluated following CIELab, using gray, black, and white backgrounds. All analyses were performed immediately after making the samples and repeated after thermal aging with 2 thousand cycles (5-55 °C). The results obtained were statistically analyzed with a significance level of 5%. FT-IR analysis showed about a 46% degree of conversion on the surface and 37% in the center of the resin sample. The flexural strength was higher for the groups polymerized for 32 min and 1h, while the Knoop microhardness did not show a statistical difference between the groups. Color and translucency analysis also did not show statistical differences between groups. According to all the analyses carried out in this study, for the evaluated material, the post-polymerization time of 1 hour can be suggested to improve the mechanical performance of 3D-printed devices.

In recent years, Additive Manufacturing (AM), commonly referred to as 3D printing, has garnered the attention of the scientific community due to its capacity to transform ordinary and traditional items into customized materials at an affordable cost, through various AM processes. Antimicrobial/antibiofilm 3D printed materials are one of the most trending research topics, owing to the growing concerns over the emergence of complex microbial structures called "biofilms" on various surfaces. The review provides an overview of the evolution of additive manufacturing (AM) technologies and their various derivatives, along with a brief description of their materials and applications. It also introduces how biofilms can represent an advantageous lifestyle for microbial populations. The primary objective of this research was to conduct a systematic review on the development of planctonic or biofilm forms of microorganisms on 3D printed materials. The article summarizes commonly studied microorganisms on these materials and presents their 3D printing process, materials, as well as the fields covered by each of the analyzed papers. To the best of our knowledge, this is the first all-inclusive systematic review that amalgamated research conducted in diverse fields to assess the development of biofilms on surfaces produced through three-dimensional printing. Most notably, this review presents a comprehensive account of sustainable approaches for producing antimicrobial materials through 3D printing. Additionally, we assess their advancements in various fields such as medicine, environment, agri-food, and other relevant sectors. The findings of our literature review can be used to recommend appropriate microorganisms, 3D printing materials, and technologies for academic and industrial research purposes focusing on the development of microbial biofilms on 3D printed surfaces. Furthermore, it highlights the potential of environmentally-friendly modified AM technologies to combat biofilms in clinical and non-clinical areas. Our goal with this review is to help readers gain a better understanding of fundamental concepts, inspire new researchers, and provide valuable insights for future empirical studies focused on eradicating biofilms from 3D printed materials.

Bursera fagaroides popularly used in México possess bioactive lignans. These compounds are low in the bark, and its extraction endangers the life of the trees. The aim of the present investigation was to search for alternative sources of cytotoxic compounds from B. fagaroides as leaves and in vitro callus cultures. The friable callus of B. fagaroides were established using a combination of phytoregulators: 4 mgL-1 of 2,4-dichlorophenoxyacetic acid (2,4-D), 1 mgL-1 Naphthaleneacetic Acid (NAA) and 1 mgL-1 Zeatin. The maximum cell growth was at day 28 with a specific growth rate μ= 0.059 days-1 and duplication time td=11.8 days. Scopoletin with a concentration of 10.7 µg/g dry weight was the main compound inducible as a phytoalexin by the addition of high concentrations of 2,4-D, as well as by the absence of nutrients in the culture medium. The com-pounds γ-sitosterol, and stigmasterol were also present. Open column chromatography was used to separate and identify: yatein, acetyl podophyllotoxin and 7'8'-dehydropodophyllotoxin in the leaves of the wild plant. Cytotoxic activity on four cancer cell lines was tested, being PC-3 prostate carcinoma (IC50 of 12.6±4.6) the most sensitive to the wild-type plant extract and HeLa cervical carcinoma (IC50 of 72±5) to the callus culture extract.

As part of a project ordered by the Ministry of Science, Technology and Innovation of Brazil, an assessment of Brazilian biological collections was conducted by the Brazilian Societies of Botany, Microbiology, Virology and Zoology to obtain data on the current state of these collections. The goal was to provide subsidies for future public policies, including financial support and prioritization policies. Herein, we present the results of the microbiological collections, which play a fundamental role in the ex situ conservation of biodiversity and support research, development and innovation. A survey was sent to public and private institutions in Brazil and answered by 168 microbiological collections from 79 different institutions. Among these, 73 comprise public research institutions and universities, demonstrating the State's importance in preserving and safeguarding Brazilian microbial diversity. The main taxonomic groups are bacteria (present in 70.24% of the collections) and fungi (52.98% of the collections) from different Brazilian ecosystems and biomes, including several type strains. Furthermore, the collections preserve microorganisms with biotechnological potential for application in environmental protection, public health, industry, and agribusiness. However, despite all these economic and biotechnological potentials, the data analysis showed serious limitations and fragilities, especially regarding physical infrastructure and human resources, and raises alerts about the risk that Brazilian collections are subjected to.

ABSTRACT: This paper provides an overview of the complex relationship between culture, gender, and business. It highlights the importance of understanding how these factors intersect and influence various aspects of the business world, including organizational practices, leadership styles, workplace dynamics, and economic outcomes. It also explores how cultural factors shape gender roles, expectations, and opportunities within the business context. Additionally, it discusses the implications of cultural influences on women's participation, advancement, and overall experiences in the business world. Understanding the interplay between culture, gender, and business is essential for fostering inclusive and diverse workplaces. Organizations that embrace cultural diversity and gender equality tend to exhibit greater creativity, innovation, and adaptability. They also benefit from increased employee satisfaction, improved decision-making processes, and enhanced financial performance. The work also examined how the values, beliefs, socialization, and team spirit of some distinct ethnic groups such as Japanese, Chinese, Jews, Igbo, and Indians have helped in the economic growth of not the groups alone but the countries where they reside. By examining these dynamics and cultures, scholars, practitioners, and policymakers can gain insights to develop effective strategies that promote gender equality, cultural sensitivity, and inclusive practices in the business world, ultimately leading to more equitable and prosperous societies.

Background: Falls in hospitalized older patients lead to increased length of stay and poorer discharge outcomes. The Comprehensive Unit-based Safety Program (CUSP) represents a quality improvement intervention for patient safety. Objective: To evaluate CUSP implementation for the prevention of falls at a teaching hospital. Method: An interdisciplinary CUSP team, received a four-hour online module and one-day face-to-face training. The intervention was developed using the WHO implementation science plan-do-study-act framework. Process evaluation was conducted using the UK Medical Research Council framework. The primary outcome measure of patient safety culture was determined using the Safety Attitude Questionnaire (SAQ), while annual fall rates were considered the secondary outcome measure. Results: Safety rounds were conducted during the two-year intervention period, each attended by five to eight HCPs, with nine to 29 patients reviewed each month. Risk factors, plans and actions were administered to individual patients, while local issues were identified and resolved through engagement of senior executives. SAQ domains increased from before (n=39) to after (n=30) the intervention of CUSP. Fall rates reduced from 11.18 per 1,000 bed days pre-implementation to zero falls post-implementation. Conclusion: Improvements in patient safety culture were observed after the implementation of CUSP alongside a reduction in fall rates.

The goal was to determine the personal, interpersonal, and sociocultural predictors of condom use among rural indigenous adolescents. Predictor variables were selected from Bandura's Social Cognitive Theory and Leininger's Transcultural Theory. The sample consisted of 419 Nahuas adolescents randomly selected from the total number of neighborhood blocks in a rural community in Puebla, Mexico. The instruments had acceptable psychometric characteristics (Cronbach alpha and validity scores). Multiple linear regression models were used. Results: 56.8% of participants were female, and 50.40 % were students. The mean age was M = 17.5 (SD = 0.97), and the majority (63%) identified as Catholic. Age at menarche/first ejaculation (β = -1.2, p = .038), attitude toward condom use (β = .13, p < .001), ethnic identity (β = .21, p < .001), and ability to negotiate condom use (β = .13, p < .003), predicted (R2 = 22.3) condom use. This study provides a basis for the integration of the cultural values of indigenous adolescents within interventions for sexual health promotion.

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.

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.

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.

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 &lt; 10-3), for samples collected with sterile gauze, TranswabTM, and dry swab, respectively. The positive culture rate was 62% and 38% (P &lt; 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.

Recently, some authors have developed a shell-less culture system for chick embryos by using a plastic lm as culture vessels. We will show that velocity of rotation of vessels in incubator and temperature have direct relations with growth of chick embryos. We also obtain a mathematical relation between velocity, temperature and rate of growth of chick embryo.

This document presents, from environmental thinking (ecosystem - culture relations), the concept of the Main Agroecological Structure of Agroecosystems (MAS, EAP, for its acronym in Spanish), considered as a dissipative cultural structure. It discusses its possible applications (resilience, production, diversity) both inside and outside the farms. The MAS can be useful in the land planning on the farms, based on the concept of potential MAS that allows the quantification of the management of internal and external corridors, including natural vegetation. At the same time, it can be useful in the context of landscape management because it shows a series of cultural relations (economic, social, symbolic and technological) hidden from the partial analysis of landscape ecology.

The main goal of this paper is to address how quickly and to what extent are international organizational cultures, brought by the world companies after the process of privatization, being implemented in a single monolithic culture. For this purpose was adopted and applied Denison model of organizational culture, which has been chosen because it emphasizes the need for balance between requirements for organization’s stability demands and its required flexibility. Considering that a different organizational culture reflects systematic change of an entire organization, this paper focuses on exploring the differences in culture dimensions among companies in domestic and foreign ownership in Serbia. A sample of 1000 employees was statistically processed. Changes in organizational culture tend to be relatively slow. The results confirm that organizational culture is a complex working environment, concerning organizational values, which represents a fundamental element of organizations. Given that the process of company ownership changes occurred fifteen years prior to the research implementation, obtained results show effects of interaction between national and organizational culture in this, relatively short, period of time. Obtained results can be generalized to countries that are passing or have recently passed a transition, and are similar in cultural characteristics.

Using the culture-work-health model, this study investigates the factors influencing the quality of life of maritime workers. This study conducted a survey of 320 maritime workers who have experience living and working on a ship for more than six months. This self-administered questionnaire included questions on organizational culture and support, self-efficacy, perceived fatigue, as well as the quality of work life. Organizational culture and self-efficacy were identified as factors affecting the quality of work life, while organizational support was found to have an indirect effect after passing through self-efficacy and perceived fatigue. The final model accounts for 63.1% of the variance in maritime workers’ quality of life. As such, this study shows that self-efficacy is important for the quality of life of maritime workers, having both direct and indirect effects. Moreover, organizational support may prove the primary intervention point for relieving perceived fatigue and enhancing self-efficacy, thus improving the quality of work life.

This research discusses the development of a software-centered EdTech (Education Technology) class model through the implementation of a course titled "3D Time Machine" at a 4-year university in Republic of Korea over two semesters. The course focused on teaching 3D modeling software, Blender, within the history department. The primary objective of offering this course was to equip students from the digital generation with the capability to manipulate digital technology effectively. Through studying historical materials and accumulating domain knowledge, students could construct their narratives from their unique perspectives. This aimed to foster their proficiency in digital technology operation, preparing them for an education environment increasingly centered around virtual worlds. As the use of virtual worlds gains prominence in educational settings, there is a growing need to incorporate curricula that prepare students to thrive as well-prepared citizens in future societies. Assessing the digital competencies of contemporary university students and designing instructional models with particular attention to their needs is becoming increasingly important. This research draws insights from interviews, conducted in both face-to-face and written formats, with students who participated in the "3D Time Machine" course. The interviews revealed valuable insights that can be actively incorporated into the development of software-centered EdTech instructional models. They reported that they expanded their perceptions as they learned how to make their ideas tangible. The course helped students overcome their fear of digital technology. In conclusion, the course effectively employs 3D modeling technology, a software-centered EdTech, as a core element in helping students develop their narratives rapidly and diversely, thereby playing a crucial role in their ability to articulate their unique perspectives.

Pure monocular 3D reconstruction is an ill-posed problem that has attracted the research community's interest due to the affordability and availability of RGB sensors. SLAM, VO, and SFM are disciplines formulated to solve the 3D reconstruction problem and estimate the camera’s ego-motion, so many methods have been proposed. However, most of these methods were not evaluated in large datasets, under various motion patterns, had not been tested under the same metrics, and most of them had not been evaluated following a taxonomy, making their comparison and selection difficult. In this research, we performed a comparison of ten publicly available SLAM and VO methods following a taxonomy, including one method for each category of the primary taxonomy, three machine learning-based methods, and two updates of the best methods, to identify the advantages and limitations of each category of the taxonomy and test if the addition of machine learning or the updates made on those methods improved them significantly. Thus, we evaluated each algorithm under the TUM-Mono benchmark and performed an inferential statistical analysis to identify significative differences through its metrics. Results determined that sparse-direct methods significantly outperformed the rest of the taxonomy, and fusing them with machine learning techniques significantly improves the performance of geometric-based methods from different perspectives.

Most common classifications for acetabular bone defects are based on radiographic two-dimension imaging, with low reliability and reproducibility. With the rise of modern processing techniques based on 3D modelling, methodologies for volumetric quantification of acetabular bone loss are available. Our study aims to describe a new methodology for quantitative assess-ment of acetabular defects, based on 3D modelling, focused on the surface analysis of the integrity of the main anatomical structures of the acetabulum represented by four corresponding sectors (posterior, superior, anterior, and medial). The defect entity is measured as the Area Increase Ra-tio (AIR) detected in all the sectors analyzed on three planes of view (frontal, sagittal and axial) compared to healthy hemipelvises. The analysis was performed on 3D-models from the CT-scan of six exemplary specimens with a unilateral pathological hemipelvis. AIR between the native and the pathological hemipelvis was calculated for each sector, for a total of 48 analysis (range, +0.93% - +171.35%). AIR of >50% were found in 22/48(45.8%) sectors and affected mostly the posterior, medial and superior sectors (20/22, 90.9%). Qualitative analysis showed consistency of the data with the morphological features of the defects. Further studies with larger samples are needed to validate the methodology and potentially develop a new classification scheme.

In the last 20 years, TADs have become one of the topics in orthodontic science, as it becomes one of the effective anchorages in moving teeth in a 3D movement to treat malocclusion and dental-alveolar orthopedic. Problems with the collusion of the roots of the teeth and trauma have hindered the use of these TADs; one of the solutions to keep TADs safe is a step-by-step innovation of procedure to make this anchorage concept using TAD safe. Making surgical guides for single-standing TADs is essential, and in practice, it is virtually possible with the advance of today's CAD/CAM and 3D printers. The content subject in the paper contains two parts: the consideration of why a surgical guide is necessary for the implantation of TADs and the making of the Surgical guide itself.

In this study, we designed a functional neuro-cardiac model to help us examine the role of neuronal regulation and confirm the importance of neural innervation techniques for cardiac tissue regeneration. A three-dimensional (3D) bioprinted neuro-cardiac scaffold composed of a mixture of gelatin-alginate and alginate-genipin-fibrin hydrogels was developed with a 2:1 ratio of AC16 cardiomyocytes (CMs) and retinoic acid differentiated SH-SY5Y neuronal cells (NCs) respectively. A unique semi-3D bioprinting approach was adopted where the CMs were mixed in the cardiac bioink and printed using an anisotropic accordion design to mimic the physiological tissue architecture in vivo. The voids in this 3D structure were methodically filled in using a NCs-gel mixture and cross-linked. Confocal fluorescent imaging using Microtubule-associated protein 2 (MAP-2) antibodies for labeling NCs, and the MyoD1 antibody for CMs revealed functional coupling between the two cell types in the final cross-linked structure. This data confirmed the development of a physiologically functional neuro-cardiac model that can be used to study neuro-cardiac modulation under physiological and pathological conditions.

In recent years, with the development of social media, people are more and more inclined to upload text, pictures and videos on the platform to express their personal emotions, thus the number of short videos is increasing and becoming the first choice for people to socialize. Unlike the traditional way, people can convey their personal emotions and opinions through media other than words, such as video images, etc. for external information. Therefore, the expression and analysis of emotions is not only through text, but also through the analysis of emotional needs in images and videos, and the research scholars have customized products for individual users. Compared with pure text content, video information can more intuitively express users' happiness, anger and sorrow, thus short video-related applications have gained more and more popularity among Internet users in recent years. However, not all short videos on social networking sites can accurately express users' emotions, and related text information can more accurately assist sentiment analysis and thus improve accuracy. However, short video sentiment analysis based on video frame images is inaccurate in some scenarios, such as when expressing tears of joy, the sentiment expressed by the user's facial expression and voice are different, which will cause errors in the analysis of sentiment. As a result, researchers began to consider multimodal sentiment analysis to reduce the impact of the above scenarios on short video sentiment analysis. This paper focuses on proposing a sentiment analysis method for short videos. We first propose a residual attention model to make full use of the information in audio to classify the emotions contained in them. Then the text information in the dataset is classified by feature extraction. The key to extract features from text information is not only to retain the semantic information of the text, but also to explore the potential emotional information in the text, so as to ensure the integrity of the text information features. The experiments show that the sentiment analysis model proposed in this paper is more superior than the baselines.

An approach employing ultrafast laser hybrid subtractive-additive microfabrication combining ablation, 3D nanolithography and welding is proposed for the realization of Lab-On-Chip (LOC) device. Single amplified Yb:KGW fs-pulsed laser source is shown to be suitable for fabricating microgrooves in glass slabs, polymerization of fine-meshes filter out of hybrid organic-inorganic photopolymer SZ2080 inside them, and, lastly, sealing the whole chip with cover glass into a single monolithic piece. The created microfluidic device proved its particle sorting function by separating 1 μm and 10 μm polystyrene spheres in a mixture. All together, this shows that fs-laser microfabrication technology is a flexible and versatile tool for the manufacturing of mesoscale multi-material LOC devices.

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 twins are gaining in popularity for simulating complex natural and urban environments. In this context, accurate segmentation of objects within 3D urban environments is of crucial importance. The aim of this project is to develop a methodology for extracting buildings from textured 3D meshes. To this end, PicassoNet-II, a semantic segmentation architecture is employed. The methodology also incorporates Markov field-based contextual analysis to assess post-segmentation features. In addition, building instantiation is performed using cluster analysis algorithms. Training this model to fit various datasets requires a large amount of annotated data, both from Quebec City, Canada, and from simulated data. Experimental results show that the use of simulated data improves segmentation accuracy, and the DBScan algorithm proves effective in extracting isolated buildings. This project paves the way for improved applications in 3D urban modeling, offering opportunities in fields ranging from urban planning to resource management. The positive results with simulated data reinforce the impact of this research on improving digital models of our ever-changing urban environments.

3D Zernike moments based on 3D Zernike polynomials have been successfully applied to the field of voxelized 3D shape retrieval and have attracted more attention in biomedical image processing. As the order of 3D Zernike moments increases, both computational efficiency and numerical accuracy decrease. Due to this phenomenon, a more efficient and stable method for computing high-order 3D Zernike moments was proposed in this study. The proposed recursive formula for computing 3D Zernike radial polynomials combines the recursive calculation of spherical harmonics to develop a voxel-based algorithm for the calculation of 3D Zernike moments. The algorithm was applied to the 3D shape Michelangelo's David with a size of 150×150×150 voxels. As compared to the method without additional acceleration, the proposed method uses a group action of order sixteen orthogonal group and saving unnecessary iterations, the factor of speed-up is 56.783±3.999 when the order of Zernike moments is between 10 and 450. The proposed method also obtained an accurate reconstructed shape with the error rate (normalized mean square error) of 0.00 (4.17×10^-3) when the reconstruction was computed for all moments up to order 450.

This paper aims to present the method of producing visual media for communicating the Asinara Island National Park plan. These products are landscape visualizations able to foster landscape preservation both from the point of view of the management of the landscape and of the citizen's involvement in the communication of the historical and environmental values of the landscape. Starting from landscape information gathering, the research has experimented with an operative method for processing different graphic representations from the same geographic database, calibrating the outputs to different audiences, their needs, objectives, and literacy skills. Three different types of products are presented: the first is a digital, dynamic and multisectoral decision-making GIS tool for park management. The second is a 3D model, aimed to the virtual fruition. The third is a map of the zoning park plan drawn to be easily readable to the non-expert public. This case study emerges as a replicable method to process the landscape information to make different visualization tools from a single geographic model, to meet the different requirements that arise from a complex landscape planning process.

New in vitro approaches for the assessment of their toxicity are crucial for creating better risk assessments given the growing use of nanoparticles (NPs) in the production of consumer goods. Humans will be exposed to nanoparticles through the inhalation route from contaminated aerosol. A promising method for evaluating the genotoxicity of NPs in vitro uses human airway epithelial cells. This study aimed to determine whether there were differences in morphology change of human bronchial/tracheal epithelial (HBTEC) cells responded to NPs in 2D and 3D spheroid models and to evaluate genotoxicity in 2D and 3D cultures after 24 hr exposure to TiO2-, Ag- and ZnO-NPs using the comet assay. Our 2D and 3D HBTEC cultures with TiO2-NPs did not show any statistically significant genotoxicity compared to controls for concentration points tested in vitro. HBTEC cells grown as 2D indicated that exposure to ZnO-NPs at low concentrations (10-50 ug/mL) for 24 hr did not cause any genotoxic effect but, at high concentrations, genotoxicity was observed. In contrast, treatment of 2D HBTEC with AgO-NPs led to an elevated in percentage of tail DNA damage from 50 ug/mL to 100 ug/mL, which was concentra-tion-dependent. Using 3D spheroid assays revealed a lower %tail length in ZnO- and AgO-NPs, thus exhibiting in vitro genotoxicity of NPs in this cell model. Spheroids (3D) rather than mono-layer cultures (2D) are believed to more precisely represent in vivo-like cell behavior when evaluating genotoxicity. Together, our findings demonstrate the genotoxic effects of NPs and point to the critical role that cell culture models play in the evaluation of toxicological risk.

In the production of green parts from powder, there is unavoidable slight deviation in the die filling, even when high-quality powders are used. The quantity of powder in the die varies and thus affects the weight of the compact. This filling variation results in variation of the pressing force, and thus influences the part geometry. The development of the DORST Netshape® System was conceived as an autonomous manufacturing system in order to compensate for these effects. Based on the Dorst Industry 4.0 innovations for part weight measuring immediately after pressing in combination with a laser dimension measuring system, this technology package attempts to reach enhanced precision and consistency in production. The paper presents results from various trials that show the capability of this new system, designed to improve the quality of pressed parts.

This paper presents the results of the investigation of composite sinters W-TiB2 which were used as an electrode in the process of electro-spark deposition (ESD) and the examination of the deposited layers. The scope of the study includes detailed characteristics of powder mixtures, composite sinters made using the spark plasma sintering method (SPS) and layers deposited in the electro-spark process. The ESD process, using the W+30 vol.% TiB2 electrode, was carried out using an automated device. The substrates were made of copper and aluminium. The topography analysis of the surfaces of the composite layer and the evaluation of their wear resistance properties are also presented. The analysis of the results of research showed the possibility to obtain, using the SPS method, composite materials of good quality, which can be used as electrodes in the ESD process. The obtained layers had increased wear resistance in relation to the substrate material.

We present progress in fast, high-resolution imaging, material classification, and fault detection using hyperspectral X-ray measurements. Classical X-ray CT approaches rely on data from many projection angles, resulting in long acquisition and reconstruction times. Additionally, conventional CT cannot distinguish between materials with similar densities. However, in additive manufacturing, the majority of materials used are known a priori. This knowledge allows to vastly reduce the data collected and increase the accuracy of fault detection. In this context, we propose an imaging method for non-destructive testing of materials based on the combination of spectral X-ray CT and discrete tomography. We explore the use of spectral X-ray attenuation models and measurements to recover the characteristic functions of materials in heterogeneous media with piece-wise uniform composition. We show by means of numerical simulation that using spectral measurements from a small number of angles, our approach can alleviate the typical deterioration of spatial resolution and the appearance of streaking artifacts.

Fatigue remains a challenge especially for high end metal AM parts and materials. From a design perspective, the fatigue limit of an AM solution can be improved upon by optimizing the manufacturing process for a certain material and part. In addition, improved accuracy of design methodologies aids in capturing the features critical for fatigue and quantifies their significance to desired part lifetime as well as providing a basis for their avoidance. In current work we present an overall concept merging thermomechanical process and powder bed solidification modeling to micromechanical analysis of fatigue of the resulting material microstructure. Material features critical to fatigue, particularly surface roughness, internal defects such as porosity and cracks and on the other hand inclusions, can be assessed directly on the basis of AM part microstructure with respect to the resulting fatigue limit. Case analyses consist of maraging steel and nickel alloys. The overall scheme provides a basis for optimization of metal AM solutions against fatigue and multiscale modeling founded basis for fatigue design.

Quality assurance has been one of the major challenges in laser-based additive manufacturing (AM) processes. This study proposes a novel process modeling methodology for layer-wise in-situ quality monitoring based on image series analysis. An image-based autoregressive (AR) model has been proposed based on the image registration function between consecutively observed thermal images. Image registration is used to extract melt pool location and orientation change between consecutive images, which contains sensing stability information. Subsequently, a Gaussian process model is used to characterize the spatial correlation within the error matrix. Finally, the extracted features from the aforementioned processes are jointly used for layer-wise quality monitoring. A case study of a thin wall fabrication by a Directed Laser Deposition (DLD) process is used to demonstrate the effectiveness of the proposed methodology.

Additive Manufacturing (AM) simplifies the fabrication of complex geometries. Its scope has rapidly expanded from the fabrication of pre-production visualization models to the manufacturing of end use parts driving the need for better part quality assurance in the additively manufactured parts. Machine learning (ML) is one of the promising techniques that can be used to achieve this goal. Current research in this field includes the use of supervised and unsupervised ML algorithms for quality control and prediction of mechanical properties of AM parts. This paper explores the applications of supervised learning algorithms - Support Vector Machines and Random Forests. Support vector machines provide high accuracy in classifying the data and is used to decide whether the final parts have the desired properties. Random Forests consist of an ensemble of decision trees capable of both classification and regression. This paper reviews the implementation of both algorithms and analyzes the research carried out on their applications in AM.

OBJECTIVE: Although 3D-printed anatomic models are not new to medicine, the high costs and lengthy production times entailed have limited their application. Our goal was developing a new and less costly 3D modeling method to depict organ-tumor relations at faster printing speeds. METHODS: We have devised a method of 3D modeling using DICOM images. Coordinates are extracted at a specified interval, connecting them to create mesh-work replicas. Adjacent constructs are depicted by density variations, showing anatomic targets (ie, tumors) in contrasting color. RESULTS: An array of organ solid-tumor models were printed via Fused Deposition Modeling 3D printer at significantly less cost ($0.05/cm3) and time expenditure (1.73 min/cm3; both, p<.001). Printed models helped promote visual appreciation of organ-tumor anatomy and adjacent tissues. Our mesh-work 3D thyroidal prototype reproduced glangular size/contour and tumor location, readily approximating the surgical specimen. CONCLUSIONS: This newly devised mesh-type 3D printing method may facilitate anatomic modeling for personalized care and improve patient awareness during informed surgical consent.

3D printing facilitates the straightforward construction of microchannels with complex three-dimensional architectures. Here, we demonstrate 3D-printed modular mixing components that operate on the basis of splitting and recombining fluid streams to decrease interstream diffusion length. These are compared to helical mixers that operate on the principle of chaotic advection.

This paper explores teachers’ professional practices to gain a thorough grasp and a deep understanding of what it takes to retain learners in an environment where most schools fail to do so. I approach the study using a qualitative multiple case study design and position it within the interpretive paradigm. I purposively selected 50 post-level one teachers from five schools. Cavanagh and Dellar’s theory on culture of school improvement, which emphasizes the transformation of schools into learning communities through social interaction, interpersonal relationships, and support among various stakeholders, including parents, teachers, and learners, was used as theoretical lens for this research. To generate data, I used focus group discussions. I present and discuss data through four themes namely, fundamental values, ethics and behaviour in the classroom, their influence in building learners’ moral vocabulary in the classroom, Classroom rituals in running the classroom smoothly and learner productivity, the effects of the celebrations on the classroom-learning communities, role served by heroes and heroines in learners learning together. In conclusion, one dynamic that stood out is that not all values that teachers had were mutually consistent or harmoniously realisable. Despite that, teachers focused on the central task of teaching, learning and management with a sense of responsibility, purpose and commitment. Teachers seemed to have mind-sets that supported a good work ethic, achievement, commitment, passion and love to attend their teaching duties diligently. Teachers uphold the values through rituals that symbolised order, that prepared students for learning and helped in binding learners of the school and teachers to each other and emphasised the unique values of the school culture. They also uphold them by celebrating academic achievement and had learners and other people who were taken as heroes and heroines of the school and were looked up to by other learners and members of the staff and generally set the tone for the way things were done.

Microfluidic systems offer precise control over physiological, biochemical, and mechanical stimuli in cell culture, allowing in vitro emulation of tissue or tumor microenvironments. This study aims to advance our understanding of tumor biology and contribute to personalized therapy development. Overcoming challenges in cell culture, including cell density and microfluidic device properties, is essential for this purpose. We designed a microfluidic system to facilitate the interaction of diverse cell lineages, incorporating human brain microvascular endothelial cells (HBEC5i), glioblastoma multiforme cells (U87MG), and astrocytes (ScienceCells 1800). Post-fabrication, we assessed the system's functionality and systematically evaluated cell inoculation conditions. Photographic acquisitions comprehensively captured cell culture and their interactions. The device successfully enabled assessment of cellular proliferation, invasion, and migration, supporting the growth of monolayers and tumor organoids. Importantly, each channel demonstrated independence in study processes while allowing cellular intercommunication, both physically and biochemically. In conclusion, the functional microfluidic device facilitated the study of cell migration, tumor invasion, and organoid growth, faithfully replicating biological features observed in a tumor microenvironment. This research contributes to advancing our understanding of tumor biology for the development of personalized therapies.

The fluctuation of the conditions on Lake Limboto affects people's activities around the lake. They try to survive by facing the problem of the lake, which is shallowing and narrowing. Yet the lake is predicted will be disappeared in 2030. This means that the productivity of the lake also diminishes. This article aims to analyze how the communities around the lake respond to the evolution and shocking prediction toward the lake into their daily activities. This study uses an interdisciplinary approach by integrating some fields of study to understand and solve the problems. One of the approaches involves interviewing people around the lake. Nevertheless, the base of the study is socio-cultural studies, which uses also old texts and oral literature as the support of the study. The research needs the theory of cultural studies to look at the survival and resilience of the people of lake so that they form their special daily activity. The results of the study show that some people know that there is a prediction of disappearing of the lake, but they have not anticipated it, as if this problem is only a government affair. They still live at the lake bank, try to survive and to be resilience even in poor conditions.

Breast cancer remains one of the most deadly non-communicable diseases in the world. The incidence of breast cancer in South Africa is increasing, with rural African women presenting with advanced stages of the disease. In this study, we aim to explore sociocultural factors influencing breast cancer screening practices among rural African women. A qualitative exploratory study was conducted using semi-structured interviews with 22 rural African women selected by purposive sampling. Thematic analysis was used to analyze the data. In this study, four sociocultural factors were identified as influencing breast cancer screening practices among rural African women. These factors included psychological factors, habits, beliefs, and healthcare perception. Women in rural African communities have deep-rooted traditional beliefs and practices regarding breast cancer. Consequently, this influences women's preventative health behaviours regarding breast cancer screening. To increase the number of women participating in breast cancer screenings, it is vital to develop culturally sensitive health education programs. Engaging community healers will also help to increase the number of women participating in breast cancer screening.

(1) Background: Microalgae cultures are greatly facilitated if growers can easily and economically ascertain the quantitative and qualitative status of the culture continuously with satisfactory ac-curacy. (2) Methods: The locally isolated microalgae Dunaliella sp. and Anabaena sp. were cultured in small volumes with 2 intensities of white light (2000 and 8000 lux) and with green, blue and red light and the increase of their biomass and pigments was studied. Pigment analyses, continuous recordings of absorption spectra and calibration curves were performed. (3) Results: The intensity of 8000 lux white light yielded the highest increase in biomass, chlorophylls and carotenoids in Dunaliella sp. while 2000 lux and green light caused the greatest increase in biomass and phy-cocyanin in Anabaena sp. From the examination of the absorption spectra, the evolution of the pigment content can be estimated and both pigments and biomass are correlated very satisfactorily with the wavelength of 750 nm. (4) Conclusions: The use of absorption spectra as an easy, fast and economical method can be a useful tool for a good approximation of the state of the microalgae culture. This is clearly shown when the spectra of the culture under different lighting colors are compared which have a catalytic effect on the level of the pigments leading to the increase in carotenoids and phycocyanin of the green light.

Lack of reliable information on PTC industry in Sri Lanka is hampering the advancement of the technology. Hence, this study attempted to assess the current status of PTC industry in Sri Lanka in order to ascertain the type and the level of interventions needed to broaden the horizons of the industry. Data of last 05 years were collected through qualitative research methods including a questionnaire-based survey, personal interviews, etc. and personal interviews to identify product diversity, R&D, available facilities, production capacity, and markets. Information was analyzed qualitatively using descriptive statistical software to assess the current status, and identify gaps, challenges and opportunities. COVID pandemic and the economic crisis had a heavy toll on the PTC industry. Six major challenges experienced by business owners were identified as increasing capacity, opportunities to build market linkages, demand fluctuations, issues relating to awareness and insufficient support given by the government. Proper identification and screening of mother plants, determining the production capacities and marketing, knowledgeable and skilled human resources, were identified as important contributory factors for success. Development strategies identified mainly include instinctive decision making and government support. Embracing the significant trade achievements through resource reallocation, prioritization and improvisation processes at individual, family group, inter-organizational levels and across these levels need to be regulated through a responsible authority. These interpretations and recommendations of this research can be utilized both by the policy makers and public and private sector organizations for decision making purposes targeting commercial scale advancements.

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).

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.

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.

Elite camels often suffer from massive injuries. Thus, there is a pivotal need for a cheap and readily available regenerative medicine source. We isolated novel stem-like cells from camel skin and investigated their multipotency and resistance against various stresses. Skin samples were isolated from ears of five camels. Fibroblasts, keratinocytes, and spheroid progenitors were extracted. After separation of different cell lines by trypsinization, all cell lines were exposed to heat shock. Then, fibroblasts and dermal cyst-forming cells were examined under cryopreservation. Dermal cyst-forming cells were evaluated for resistance against osmotic pressure. The results revealed that resistance periods against trypsin were 1.5, 4, and 7 minutes for fibroblasts, keratinocytes, and spheroid progenitors, respectively. Furthermore, complete recovery of different cell lines after heat shock along with the differentiation of spheroid progenitors into neurons was observed. Fibroblasts and spheroid progenitors retained cell proliferation after cryopreservation. Dermal cyst-forming cells regained their normal structure after collapsing by osmotic pressure. The spheroid progenitors incubated in the adipogenic, osteogenic, and neurogenic media differentiated into the adipocytes, osteoblasts, and neurons, respectively. To the best of our knowledge, we isolated different unique cellular differons and stem-like cells from the camel skin and examined their multipotency for the first time.

Building an ethical culture is challenging and a basic requirement of the companies to embed transparency in their systems, creating a positive image; serving the internal and external communities. This paper based on detailed interviews of 12 family owned businesses over a period of 16 months explores how these companies build the ethical culture, identifying the ethical culture sustainability triggers, challenges and role of religion in such practices. The values, culture, community and social norms are identified as major ingredients of a sustainable ethical culture development and implementation of the ethical policies and procedures require institutional and structural mechanisms for effectiveness in family owned businesses. The findings at numerous occasions are in contrast to the literature, whereas, in other instances are similar. The religion, society, family image, the entrepreneurs themselves and their family members play a vital role. Non-interference in private matters of the employees, whistle blowing, code of ethics, training and awareness creation and a number of other factors play a leading role in ethical culture development in family businesses.

Abstract Sustainability of ethnic culture in Southeast Asia has made the dramatically growth of ethnic identity. The ethnic revivals already made the increasing of cultural events in public spaces. This research paper sought the cultural sustainability of Chinese in Moslem society of Southaest Asia. A multisited ethnography was conducted in Medan Municipal of North Sumatra and Georgetown Penang, Malaysia to observe the sustained Chinese culture as the symbol of ethnification of Chinese in Moslems society in Southeast Asia region. It found that after 2003 Indonesia already saw the attractive cultural performances of Chinese in public spaces as the continuation of sustainability. In our ethnographic investigation from 2014 -2017, the reshaping of the Chinese identity through sustainability of Chinese culture in Medan Municipality of North Sumatra, and Penang of Malaysia has the high public visibility. Research report showed the continuation of the Chinese rituals and festivals which were accompanied by music instruments of Chinese and theatrical performances. Those have been transformed from self commemorations to be more public; attractions already were moved to public places, not solely in temples or ethnic group surroundings as what commonly found in the past period. It concluded that the sustainability of Chinese culture in public spaces made the Southeast Asia connection among the Chinese groups solidify their identity in this region tightened.

The current discussion about a “risk culture” in financial services was triggered by the recent series of crisis (from the US subprime mortgage crisis to the lock-in trap of central banks in the aftermath of the sovereign debt crisis). The last decade saw a long list of hybris, misconduct and criminal activities by human beings on a single or even a collective basis in banks, in the industry or in the whole economy. As a counter-reaction, financial authorities called for a guidance by a “new” risk culture in financial institutions based on a set of abstract, formal and normative governance processes. In the humanities, culture is a paraphrase for the behaviour in collectives and dynamics of organization found in human societies. Therefore, a “risk culture” should be a link between those normative guidelines and the positive “real-world” decision-making in financial services. This paper will focus on “risk culture” from the perspective of human beings interacting in dynamical and intertemporal commercial relations. In this context, “risk” is perceived by economic agents ex-ante as the consequence of the time-lag between the present and the uncertain future development (compared to a probability distribution calculated by observers ex-post). For all those individual decisions – to be made under uncertainty – future “risk” includes the so-called “normal accidents”, i.e. failures that will happen at some uncertain point in time but inevitable, and it is only the questions, when failure will happen and how to maintain function in the first line of defence. Finally, the sum of all decisions with individual responsibility for things to come is the positive economical context of a “risk culture” in the sense of the (rational) behaviour of an honourable merchant in long-term and repeated commercial relations in a merchant community, as this is a heritage of European business culture.

Tiny by physical size, the State of Israel retains some of the world’s most important cultural treasures, along with many other great cultural institutions. Archeological treasures have yielded much information as far as biblical history and have been well adapted to a Zionist narrative by both the Jewish press and international news organizations, such as the New York Times whose archives are replete with reports of Jewish history being dug up by the Jewish people. Once the State of Israel gained independence in 1948, the course was set for the development of historical museums whose discourse would reflect the most significant events in Jewish history, most especially the Holocaust and the state of constant warfare that continues to imbue the cultural consciousness of its citizens. In this paper we outline, through categorization, the various historical museums, which are currently operating. Furthermore, this article hopes to shed some light upon the cultural sensibilities conveyed through these institutions.

It is time-consuming for acquiring complete data by fully phase encoding in two orthogonal directions along with one frequency encoding direction. Undersampling in the 3D k-space is promising in accelerating such 3D MRI process. Though 3D undersampling can be conducted according to predefined probability density, the density based method is not optimal. Because of the large amount of 3D data and computational cost, it is challenging to perform data-driven and learning-based 3D undersampling and subsequent 3D reconstruction. To tackle this challenge, this paper proposes a deep neural network called EEUR-Net, realized by optimizing specific undersampling patterns for the fully-sampled 3D k-space data. Innovatively, our undersampling algorithm employs an end-to-end deep learning approach to optimize phase encoding patterns and uses a 3D U-Net for image reconstruction of undersampled data. Through end-to-end training, we obtain an optimized 3D undersampling pattern, which significantly enhances the quality of the reconstructed image under the same acceleration factor. A series of experiments on a knee MRI dataset demonstrated that, in comparison to standard random uniform, radial, Poisson and equispaced Cartesian undersampling schemes, our end-to-end learned undersampling pattern considerably improves the reconstruction quality of undersampled MRI images.

Three-dimensional reconstruction plays an important role in assisting doctors and surgeons in diagnosing bone defects’ healing progress. Common three-dimensional reconstruction methods include surface and volume rendering. As the focus is on the shape of the bone, volume rendering is omitted. Many improvements have been made on surface rendering methods like Marching Cubes and Marching Tetrahedra, but not many on working towards real-time or near real-time surface rendering for large medical images, and studying the effects of different parameter settings for the improvements. Hence, in this study, an attempt towards near real-time surface rendering for large medical images is made. Different parameter values are experimented on to study their effect on reconstruction accuracy, reconstruction and rendering time, and the number of vertices and faces. The proposed improvement involving three-dimensional data smoothing with convolution kernel Gaussian size 0.5 and mesh simplification reduction factor of 0.1, is the best parameter value combination for achieving a good balance between high reconstruction accuracy, low total execution time, and a low number of vertices and faces. It has successfully increased the reconstruction accuracy by 0.0235%, decreased the total execution time by 69.81%, and decreased the number of vertices and faces by 86.57% and 86.61% respectively.

Solar3D is an open-source software application designed to interactively calculate solar irradiation at three-dimensional (3D) surfaces in a virtual environment constructed with combinations of 3D city models, digital elevation models (DEMs), digital surface models (DSMs) and feature layers. The GRASS GIS r.sun solar radiation model computes solar irradiation based on two-dimensional (2D) raster maps for given day, latitude, surface and atmospheric conditions. With the increasing availability of 3D city models and demand for solar energy, there is an urgent need for better tools to computes solar radiation directly with 3D city models. Solar3D extends GRASS GIS r.sun from 2D to 3D by feeding the model with input, including surface slope, aspect and time-resolved shading, that is derived directly from the 3D scene using computer graphics techniques. To summarize, Solar3D offers several new features which, as a whole, distinguish itself from existing 3D solar irradiation tools: (1) the ability to consume massive heterogeneous 3D city models, including massive 3D city models such as oblique airborne photogrammetry-based 3D city models (OAP3Ds or integrated meshes); (2) the ability to perform near real-time pointwise calculation for duration from daily to annual; (3) the ability to integrate and interactively explore large-scale heterogeneous geospatial data. (4) the ability to calculate solar irradiation at arbitrary surface positions including at rooftops, facades and the ground. Solar3D is publicly available at https://github.com/jian9695/Solar3D.

3D printing is described as the third industrial revolution: its impact is global in industry and progresses every day in society. It presents a huge potential for ecology and evolution, sciences with a long tradition of inventing and creating objects for research, education and outreach. Its general principle as an additive manufacturing technique is relatively easy to understand: objects are created by adding material layers on top of each other. Although this may seem very straightforward on paper, it is much harder in the real world. Specific knowledge is indeed needed to successfully turn an idea into a real object, because of technical choices and limitations at each step of the implementation. This article aims at helping scientists to jump in the 3D printing revolution, by offering a hands-on guide to current 3D printing technology. We first give a brief overview of uses of 3D printing in ecology and evolution, then review the whole process of object creation, split into three steps: (1) obtaining the digital 3D model of the object of interest, (2) choosing the 3D printing technology and material best adapted to the requirements of its intended use, (3) pre- and post-processing the 3D object. We compare the main technologies available and their pros and cons according to the features and the use of the object to be printed. We give specific and key details in appendices, based on examples in ecology and evolution.

Excess consumption of a high-fat diet has been identified as one of the contributing factors for obesity. Women are at higher risk for adult obesity than men, predisposing them to a different set of detrimental disease conditions. Furthermore, aging studies show that physiological decline also has a serious impact on changes in the endocrine properties of white adipose tissue. Howev-er, there is still relatively little known about the factors associated with obesity and aging and their compounding impact on women’s health. To investigate adipocytokine secretion profile changes, female C3H/HeJ mice were induced to obesity through long-term consumption of a high-fat diet. Weight gain measurements and Echo MRI analysis of fat composition showed in-creases were a due solely to the high fat content in the diet. Adipocytokine secretions were ana-lyzed in media conditioned from harvested visceral fat tissue that was organ cultured ex-vivo. Adipocytokine analysis across diets and age showed significant increases in Adiponectin and Leptin secretion in high-fat diets, accelerated increases in Resistin secretion with high-fat diets. Aging induced increased secretion of Lipcalin-2, Pentraxin-3 and Serpin E1, MCP-1 and ICAM-1, regardless of diet. Furthermore, comparisons of organoid cultured protein secretions and flash-frozen tissue samples differed greatly, suggesting the WAT organoid cultures may yield information more reflective of in situ conditions. Taken together, our results show that diet and aging significantly impact secretions from adipose tissue, that may contribute to women’s health issues.

Cellular agriculture is defined as the production of agricultural products from cell cultures rather than from whole plants or animals. With growing interest in cellular agriculture as a means to address the public health, environmental, and animal welfare challenges of animal agriculture, the concept of producing seafood from fish cell- and tissue-cultures is emerging as a means to address similar challenges with industrial aquaculture systems and marine capture. Cell-based seafood - as opposed to animal-based seafood - can combine developments in biomedical engineering with modern aquaculture techniques. Biomedical engineering developments such as closed-system bioreactor production of land animal cells create a basis for large scale production of marine animal cells. Aquaculture techniques such as genetic modification and closed system aquaculture have achieved marked gains in production that can pave the way for innovations in cell-based seafood production. Here, we present the current state of innovation relevant to the development of cell-based seafood across multiple species as well as specific opportunities and challenges that exist for advancing this science. The authors find that the physiological properties of fish cell- and tissue- culture may be uniquely suited to cultivation in vitro. These physiological properties, including hypoxia tolerance, high buffering capacity, and low-temperature growth conditions, make marine cell culture an attractive opportunity for scale production of cell-based seafood; perhaps even more so than mammalian and avian cell cultures for cell-based meats. This, coupled with the unique capabilities of crustacean tissue-friendly scaffolding such as chitosan, a common seafood waste product and mushroom derivative, presents great promise for cell-based seafood production via bioreactor cultivation. To become fully realized, cell-based seafood research will require more understanding of fish muscle culture and cultivation; more investigation into serum-free media formulations optimized for fish cell culture; and bioreactor designs tuned to the needs of fish cells for large scale production.

We introduce optically clear and resilient free-form micro-optical of pure (non-photosensitized) organic-inorganic SZ2080 material made by femtosecond 3D laser lithography (3DLL). This is advantageous for rapid printing of 3D micro-/nanooptics, including their integration directly onto optical fibers. A systematic study on the fabrication peculiarities and quality of resultant structures is performed. Comparison of microlenses’ resiliency to CW and femtosecond pulsed exposure is determined. Experimental results prove that pure SZ2080 is ∼3 fold more resistant to high irradiance as compared with a standard photo-sensitized material and can sustain up to 1.91 GW/cm² intensity. 3DLL is a promising manufacturing approach for high-intensity micro-optics for emerging fields in astro-photonics and atto-second pulse generation. Additionally, pyrolysis is employed to shrink structures up to 40% by removing organic SZ2080 constituents. This opens a promising route towards downscaling photonic lattices and creation of mechanically robust glass-ceramic structures.

Aerococcus urinae is an aerobic Gram-positive coccus that grows as tiny alpha-hemolytic colonies. Actinotignum schaalii is a slow-growing facultative anaerobic Gram-positive rod. These bacteria are part of the urogenital microbiota of healthy patients, but can also be involved in urinary tract infections (UTIs), particularly in elderly men and young children [1–3]. Because A. urinae and A. schaalii are fastidious and are difficult to identify with phenotypic methods, they are underestimated causes of UTIs. Their growth is slow and requires a blood-enriched medium and an incubation time of 48 hours under anaerobic or 5% CO2 atmosphere. Furthermore, accurate identification is only possible using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) or molecular-based methods. In rare cases, these bacteria can be responsible for invasive infections. We describe here the second case of bacteremic UTI caused by both A. schaalii and A. urinae in an 89-year-old woman who presented dyspnea and asymptomatic bacteriuria. This challenging clinical and microbiological diagnosis was made by our laboratory by Gram staining urine with a leucocyte count &gt;50/μL and/or a bacterial count &gt;14/μL, urinary culture on blood agar plate, and direct bacterial identification on positive blood culture bottles (BCBs) by MALDI-TOF MS. After 10 days of antimicrobial treatment consisting of 2g amoxicillin PO t.i.d the patient was discharged with complete clinical and biological recovery. A. schaalii and A. urinae are probably still underestimated causes of UTIs. Microbiologists should assess the presence of these two bacteria in urine using the appropriate culture and identification methods where direct examination is positive for small Gram-positive cocci or rods, and a urinary dipstick is negative for nitrites and associated with leukocyturia.