Although conventional immunohistochemistry for neurotropic Rabies virus (RABV) usually shows a high preference for neurons, non-neuronal cells are also potential target cells and abortive infection of astrocytes is considered a main trigger of innate immunity in the CNS. While in vitro studies indicated differences between field and less virulent lab-adapted RABVs, a systematic and quantitative comparison of astrocyte tropism in vivo is lacking. Here, a recently developed solvent-based tissue clearing technique was used to measure the RABV cell tropism in infected brains. Immunofluorescence analysis of 1 mm-thick tissue slices enabled 3D segmentation and quantification of infection frequencies of astrocytes and neurons. Comparison of highly virulent street virus clones from fox, dog, and raccoon with three lab strains of intermediate and low virulence revealed remarkable differences in the ability to infect astrocytes in vivo. While all viruses and infection routes led to comparable neuron infection frequencies, striking differences were detected for the infection of astrocytes. Consistent and inoculation route-independent astrocyte infection by field viruses, together with route-dependent or undetectable astrocyte infection by lab-adapted or vaccine viruses strongly suggests a model in which the ability to establish productive astrocyte infection in vivo functionally distinguishes field and attenuated lab RABV strains.

Duplex Stainless Steels (DSS) and Superduplex Stainless Steels (SDSS) are an important class of stainless steels because they combine the benefits of austenite and ferrite phases, resulting in steels with better mechanical properties and higher corrosion resistance. Due to these characteristics are widely employed in various industries. However, the appearance of deleterious phases in their microstructure impairs the properties of DSS and SDSS. Among the deleterious phases, the main one is the sigma phase (σ), which can be nucleated when the steel is exposed to the temperature range between 650 °C and 900 °C, reducing its toughness and resistance to corrosion. In a previous work, Fonseca and collaborators used two descriptors of the microstructural path to analyze the formation of sigma phase (σ), SV, interfacial area per unit volume between sigma phase and austenite, and <λ>, mean chord length of sigma, both in function of the VV, volume fraction of sigma, known in the literature as microstructural partial path (MP). In this work, the contiguity ratio is applied for the first time to describe the microstructural path in the study of sigma phase precipitation in SDSS. The contiguity ratio showed that the distribution of the ferrite/sigma boundaries is homogeneous. Thus, it is reasonable to infer that one has a uniform distribution of sigma phase nuclei within the ferrite. About the kinetics of sigma phase formation, the DSS can be described by the classical JMAK equation, whereas for the SDSS, the kinetics tends to follow the Cahn model for grain edge nucleation. Finally, we present the 3D reconstruction of the sigma phase in SDSS. The results demonstrate that the sigma phase nucleates at the edges of the ferrite/austenite interfaces. Moreover, the sigma phase grows consuming the ferrite, but it is not fully interconnected.

Buildability, i.e. the ability of a deposited material bulk to retain its dimmensions under increasing load, is an inherent prerequisite for formwork-free digital construction (DC). Since DC processes are relatively new, no standard methods of characterization are available yet. The paper at hand presents practice-oriented buildabilty criteria by taking various process parameters and construction costs into consideration. In doing so, direct links between laboratory buildability tests and target applications are established. A systematic basis for calculating the time interval (TI) to be followed during laboratory testing is proposed for the full-width printing (FWP) and filament printing (FP) processes. The proposed approach is validated by applying it to a high-strength, printable, fine-grained concrete. Comparative analyses of FWP and FP revealed that to test the buildability of a material for FP processes, higher velocities of the printhead should be established for laboratory tests in comparison to those needed for FWP process, providing for equal construction rates.

3D bioprinting holds great promise in the field of regenerative medicine as it can create complex structures in a layer-by-layer manner using cell-laden bioinks, making it possible to imitate native tissues. Current bioinks lack both the high printability and the biocompatibility required in this respect. Hence, the development of bioinks that are capable of both properties is needed. In our previous study, a furfuryl-gelatin based bioink, crosslinkable by visible light, was used for creating mouse mesenchymal stem cell-laden structures with high fidelity. In this study, lattice mesh geometries were printed in a comparative study to test against the properties of a traditional rectangular-sheet. After 3D printing and crosslinking, both structures were analysed for swelling and rheological properties, and their porosity estimated using scanning electron microscopy. Results showed that the lattice structure was relatively more porous but sturdy and exhibited a lower degradation rate compared to the rectangular-sheet. Further, the lattice allowed encapsulation of a greater number of cells, allowing them to proliferate to a greater extent compared to the rectangular-sheet that retained a lesser number of cells initially. All of these results collectively affirmed that the lattice poses as a superior scaffold design for tissue engineering applications.

3D plotting is an additive manufacturing technology enabling biofabrication, thus the integration of cells or biologically sensitive proteins or growth factors into the manufacturing process. However, most (bio-)inks developed for 3D plotting were not shown to be processed into clinical relevant geometries comprising critical overhangs and cavities, which would collapse without a sufficient support material. Herein, we have developed a support hydrogel ink based on methylcellulose (mc), which is able to act as support as long as the co-plotted main structure is not stable. Therefore, 6 w/v %, 8 w/v % and 10 w/v % mc were allowed to swell in water, resulting in viscous inks, which were characterized for their rheological and extrusion properties. The successful usage of 10 w/v % mc as support ink was proven by multichannel plotting of the support together with a plottable calcium phosphate cement (CPC) acting as main structure. CPC scaffolds displaying critical overhangs or a large central cavity could be plotted accurately with the newly developed mc support ink. The dissolution properties of mc allowed complete removal of the gel without residuals, once CPC setting was finished. Finally, we fabricated a scaphoid bone model by computed tomography data acquisition and co-extrusion of CPC and the mc support hydrogel.

A novel and simple method to improve the corrosion resistance of copper by constructing a 3D 1-dodecanethiol self-assembled monolayers (SAMs) in 3.5% NaCl solution is reported in this study. Several drops of 1% H3PO4 solution are thinly and uniformly distributed on copper surface to form a 3D nanostructure constituted by Cu3(PO4)2 nanoflowers. The anticorrosion properties of 1-dodecanethiol SAMs on copper surface and on copper surface treated with H3PO4 solution were evaluated. Results demonstrated that 1-dodecanethiol SAMs on bare copper surface exhibit good protection capacity, whereas a copper surface pretreated with H3PO4 solution can substantially enhance the corrosion resistance of 1-dodecanethiol SAMs.

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.

Inexpensive piezoelectric diaphragms can be used as sensors to facilitate both nozzle height setting and build platform leveling in FFF (Fused Filament Fabrication) 3D printers. Tests simulating nozzle contact are conducted to establish the available output and an output of greater than 8 Volts found at 20 ºC, a value which is readily detectable by simple electronic circuits. Tests are also conducted at a temperature of 80 ºC and, despite a reduction of greater than 80% in output voltage, this is still detectable. The reliability of piezoelectric diaphragms is investigated by mechanically stressing samples over 100,000 cycles at both 20 ºC and 80 ºC and little loss of output over the test duration is found. The development of a nozzle contact sensor using a single piezoelectric diaphragm is described.

Wild-type p53 cancer therapy-induced senescent cells frequently engulf and degrade neighboring ones inside a massive vacuole in their cytoplasm. After clearance of the internalized cell, the vacuole persists, seemingly empty, for several hours. Despite large vacuoles being associated with cell death, this process is known to confer a survival advantage to cancer engulfing cells, leading to therapy resistance and tumor relapse. Previous attempts to resolve the vacuolar structure and visualize their content using dyes were unsatisfying for lack of known targets and ineffective dye penetration and/or retain. Here, we overcame this problem by applying optical diffraction tomography and Raman spectroscopy to MCF7 doxorubicin-induced engulfing cells. We demonstrated a real ability of cell tomography and Raman to phenotype complex microstructures, such as cell-in-cells and vacuoles, and detect chemical species in extremely low concentrations within live cells in a completely label-free fashion. We show that vacuoles had a density indistinguishable to the medium, but were not empty, instead contained diluted cell-derived macromolecules, and we could discern vacuoles from medium and cells using their Raman fingerprint. Our approach is useful for the non-invasive investigation of senescent engulfing (and other peculiar) cells in unperturbed conditions, crucial for a better understanding of complex biological processes.

Natural killer (NK) cell therapy is one of the most promising treatments for Glioblastoma Multiforme (GBM). However, this emerging technology is limited by the availability of sufficient numbers of fully functional cells. Here, we investigated the efficacy of NK cells that were expanded and treated by interleukin-2 (IL-2) and heat shock protein70 (HSP70), both in vitro and in vivo. Proliferation and cytotoxicity assays were used to assess the functionality of NK cells in vitro, after which treated and naïve NK cells were administrated intra-cranially and systemically to compare the potential antitumor activities in our in vivo rat GBM models. In vitro assays provided strong evidence of NK cell efficacy against C6 tumor cells. In vivo tracking of NK cells showed efficient homing around and within the tumor site. Furthermore, significant amelioration of the tumor in rats treated with HSP70/Il-2 treated NK cells as compared to those subjected to non-treated NK cells, as confirmed by MRI, proved the efficacy of adoptive NK cell therapy. Moreover, results obtained with systemic injection confirmed migration of activated NK cells over the blood brain barrier and subsequent targeting of GBM tumor cells. Our data suggest that administration of HSP70/Il-2 treated NK cells may be a promising therapeutic approach to be considered in the treatment of GBM.

Metal-supported fuel cells (MSCs) offer potential material cost and robustness advantage over anode supported cells (ASCs). Because of the very good thermal shock stability of such MSCs, these can be heated up very quickly. However the challenge co-firing of metal substrate and electrolyte stays still unsolved. The production of the core component, the tape casted metal substrate, with defined shrinkage resulting from modification of metal powders is presented in the paper. This approach leads to an adjustment of the shrinkage mismatch during sintering, between metal and ceramic components, and adjustment of target specifications like porosity, green density and layer thickness.

The emergence of new methodologies and tools for digitizing objects belonging to the Cultural Heritage (CH) changed the paradigms adopted so far. Traditionally, the process to acquire artifacts required specialized and often costly equipment tailored to specific purposes. However, the development of more generalized, adaptable, and affordable tools led to novel approaches. This manuscript provides a glimpse into the evolving landscape of custom-made tools for digital documentation, both hardware and software, and their transformative impact on the digitization techniques, built to meet requirements of specific case studies, including ancient drawings, manuscripts, paintings, and museum objects. The advent of self-built instruments has revolutionized the way professionals work today, by leveraging a new generation of low-cost, adaptable gears, leading to unprecedented flexibility and efficiency, while facilitating the capture of high-quality digital representations of objects with minimal damage and preserving their integrity. The outcomes of instruments and tools specifically produced for the contexts described in this paper highlight their potential for promoting interdisciplinary collaboration, facilitating scholarly research, enhancing conservation efforts, and fostering cultural exchange. Ultimately, this research contributes to illustrate how custom software, in combination with recent devices such as smartphones and 3D printers, underscores the importance of adopting these innovative approaches to generate an ecosystem of tools and methods able to preserve, document, and promote the richness of our collective past for future generations.

Antigen-presenting cells (APCs) are essential for modulating immune responses, and optimizing their differentiation protocols is critical for cell-based immunotherapies. This study compares eight different protocols for differentiating APCs from C57BL/6J mice using GM-CSF and L-929 supernatant. Four groups treated with GM-CSF and four with L-929 supernatant at various concentrations, alongside a control group were evaluated. Flow cytometry was used to analyze the expression of key markers, including MHC II, CD11c, F4/80, and CD11b. Our findings show that GM-CSF resulted in superior differentiation outcomes compared to L-929 supernatant, with higher expression levels of the analyzed markers. This study underscores the importance of considering both cell yield and protein expression quality in selecting differentiation protocols. For researchers developing therapies targeting specific cell populations, these findings demonstrate how different culture conditions influences both the quantity and phenotypic characteristics of differentiated cells. Balancing efficiency and phenotypic fidelity is crucial for optimizing cell-based immunotherapy strategies. Our data provide a valuable reference for future studies and applications involving APC differentiation from bone marrow precursors, ensuring robust foundations for developing advanced immunotherapies. This comprehensive comparison of differentiation protocols offers insights essential for enhancing the effectiveness and cost-efficiency of cell-based therapies.

The main focus of the in vitro toxicity assessment methods is on the viability of the cells, which is usually based on metabolism changes. Yet, when exposed to toxic substances, the cell triggers multiple signals in response. With this in mind, we have developed a promising cell-based toxicity method that observes various cell responses when exposed to toxic substances (either death, division or remaining viable). Based on the collective cell response, we observed and predicted the dynamics of the cell population to determine the toxicity of the toxicant. The method was tested with two different conformations: In the first conformation, we exposed a monoculture model of blood macrophages to UV light, hydrogen peroxide, nutrient deprivation, tetrabromobisphenol A, fatty acids and 5-fluorouracil. In the second, we exposed a coculture liver model consisting of hepatocytes, hepatic stellate cells, Kupffer cells and liver sinusoidal endothelial cells to rifampicin, ibuprofen and 5-fluorouracil. The method showed good accuracy compared to established toxicity assessment methods. In addition, this approach provided more representative information on the toxic effects of the compounds as it considers the different cellular responses induced by toxic agents.

Macro-autophagy (autophagy hereafter) is an evolutionarily conserved cellular process that has long been recognized as an intracellular mechanism for maintaining cellular homeostasis. It involves the formation of a membraned structure called the autophagosome, which carries cargo that includes toxic protein aggregates and dysfunctional organelles to the lysosome for degradation and recycling. Autophagy is primarily considered and studied as a cell-autonomous mechanism. However, recent studies have illuminated an underappreciated facet of autophagy, i.e., non-autonomously regulated autophagy. Non- autonomously regulated autophagy involves the degradation of autophagic components, including organelles, cargo, and signaling molecules, and is induced in neighboring cells by signals from primary adjacent or distant cells/tissues/organs. This review provides insight into the complex molecular mechanisms governing non-autonomously regulated autophagy, highlighting the dynamic interplay between cells within tissue/organ or distinct cell types in different tissues/organs. Emphasis is placed on modes of intercellular communication that include secreted molecules, including microRNAs, and their regulatory roles in orchestrating this phenomenon. Furthermore, we explore the multidimensional roles of non-autonomously regulated autophagy in various physiological contexts, spanning tissue development and aging, as well as its importance in diverse pathological conditions, including cancer and neurodegeneration. By studying the complexities of non-autonomously regulated autophagy, we hope to gain insights into the sophisticated intercellular dynamics within multicellular organisms, including mammals. These studies will uncover novel avenues for therapeutic intervention to modulate intercellular autophagic pathways in altered human physiology.

Combined radiation with hemorrhage (combined injury, CI), exacerbates hematopoietic acute radiation syndrome and mortality compared to radiation alone (RI). We evaluated the effects of RI or CI on blood cell depletion as a biomarker to differentiate the two. Male CD2F1 mice were exposed to 8.75 Gy γ- radiation (60Co). Within 2 h of RI, animals were bled under anesthesia 0% (RI) or 20% (CI) of total blood volume. Blood samples were collected at 4-5 h and days 1, 2, 3, 7, and 15 after RI. CI decreased WBC at 4-5 h and continued to decrease them until day 3; counts then stayed at the nadir up to day 15. CI decreased neutrophils, lymphocytes, monocytes, eosinophils and basophils more than RI on day 1 or day 2. CI decreased RBCs, hemoglobin and hematocrit on days 7 and 15 compared to RI, whereas Hemorrhage alone returned to the baseline on days 7 and 15. RBCs depleted after CI faster than post RI. Hemorrhage alone increased platelet counts on days 2, 3, and 7, which returned to the baseline on day 15. Our data suggest that WBC depletion may be a biomarker within 2 days post-CI and RBC depletion after 3 days post-CI.

Cell junction that is usually associated with dynamic cytoskeletons is essential for a wide range of cellular activities such as cell migration, cell communication, barrier function and signal transduction. Real-time observation of cell junction facilitates understanding of the mechanisms by which cell junction regulate relevant cellular activities. In this study, we examined the binding capacity of a modified tridecapeptide from Cx43 to cell junction protein ZO-1 and sought to create a fluorescent peptide that could be used to label cell junction. To introduce the fluorescent peptide into the cells, a cell-penetrating peptide was linked to the modified tridecapeptide and synthesized. The binding of the modified tridecapeptide was tested using pull down and immunoprecipitation assay. The ability of the peptide to label cell junction was assessed by adding it to fixed or cultured Caco-2 cells. Testing assay revealed that the Cx43-derived peptide can bind to ZO-1. Besides, the peptide was able to label cell junction of fixed cell, although no obvious cell junction labeling was observed clearly in living cells, probably due to the inadequate affinity. These results indicate that a peptide-based strategy for labeling cell junction may be feasible, and efforts to improve its affinity are warranted in the future.

The multidrug-resistant fungal pathogens belonging to the Candida haemulonii complex and the phylogenetically related species Candida auris are well-known for causing infections that are difficult to treat due to their multidrug-resistance profile. C. auris is even more worrisome due to its ability to cause outbreaks in healthcare settings. These emerging yeasts produce a wide range of virulence factors that facilitate the development of the infectious process. In recent years, the aggregative phenotype has been receiving attention, as it is mainly associated with defects in cellular division and its possible involvement in helping the fungus to escape from host immune responses. In this study, we initially investigated the aggregation ability of 18 clinical isolates belonging to the C. haemulonii species complex and C. auris. Subsequently, we evaluated the effects of physicochemical factors on the fungal aggregation competence. The results demonstrated that cell aggregation was a typically time-dependent event, in which almost all studied fungal isolates exhibited high aggregation after 2 h of incubation at 37°C. The aggregation was not impacted by pH, temperature, -mercaptoethanol (a protein-denaturing agent) and EDTA (a chelator agent). Conversely, proteinase K, trypsin and sodium dodecyl sulfate significantly diminished the aggregation. Collectively, our results demonstrated that the aggregation ability in these opportunistic yeast pathogens is time-dependent, and surface proteins and hydrophobic interactions seem to mediate cell aggregation, since the presence of proteases and anionic detergent affected the aggregation ability. However, further studies are necessary to better elucidate this phenomenon.

Different studies corroborate a role for ceramide synthases and their downstream products, ceramides, in modulation of apoptosis and autophagy in the context of cancer. These mechanisms of regulation, however, appear to be context dependent in terms of ceramides’ fatty acid chain length, subcellular localization, and the presence or absence of their downstream targets. Our current understanding of the role of ceramide synthases and ceramides in regulation of apoptosis and autophagy could be harnessed to pioneer the development of new treatments to activate or inhibit a single type of ceramide synthase, thereby regulating the apoptosis induction or cross talk of apoptosis and autophagy in cancer cells. Moreover, the apoptotic function of ceramide suggests that ceramide analogues can pave the way for the development of novel cancer treatments. Therefore, in the current review paper we discuss the impact of ceramide synthases and ceramides in regulation of apoptosis and autophagy in context of different types of cancers. We also briefly introduce the latest methods to analyze the lipids in biological samples. Finally, we discuss the drug development strategies focusing on the ceramide synthases and ceramides as future therapeutic approaches in cancer therapy.

Apple (Malus x domestica) fruit size is dependent on cell division and cell expansion, processes which are subsequently regulated by plant hormones such as auxins, gibberellins, and cytokinins. In this study, we investigated the role of cell division and cell expansion in apple growth and identified which of the two was more deterministic to final fruit size. Three cultivars of different sizes were selected, namely, ‘Twenty Ounce’ (large-sized), ‘Royal Gala’ (medium-sized), and ‘Crabapple’ (small-sized). Gene expression and cell size analyses were conducted over the course of two consecutive seasons. The expression patterns of three classes of genes were markedly similar across all cultivars. Two cell division markers, namely, MdCDKB2;2 and MdANT2, were discovered to be correlatively expressed as both displayed initially high expression levels, which gradually declined from the early to late stages of growth time course. For cell expansion markers, MdEXP3 was upregulated as the cells expanded, while MdARF106 was expressed in both the cell division and expansion stages. Meanwhile, the ripening related gene MdACO1 was expectedly expressed only during the ending stages associated with ripening. Interestingly, the cell measurements taken regularly from each cultivar throughout the same eperimental timespan showed that cell sizes were unaltered and remained constant from initial pollination at the zeroth Days After Pollination (DAP), to ripening at 120 Days After Full Bloom (DAFB).

The fate of a viral infection in the host begins with various types of cellular responses, such as abortive, productive, latent, and destructive infections. Apoptosis, necroptosis, and pyroptosis are the three major types of regulated cell death mechanisms that play critical roles in viral infection response. Cell shrinkage, nuclear condensation, bleb formation, and retained membrane integrity are all signs of osmotic imbalance-driven cytoplasmic swelling and early membrane damage in necroptosis and pyroptosis. Caspase-driven apoptotic cell demise is considered in many circum-stances as an anti-inflammatory, and some pathogens hijack the cell death signaling routes to initiate a targeted attack against the host. In this review, we selected mechanisms by which viruses interfere with cell death are discussed in-depth and are used to illustrate the general principles and cellular signaling mechanisms of virus-host specific molecule interactions.

Neural stem cell (NSC) based therapies are at the forefront of regenerative medicine strategies to combat illness and injury of the central nervous system (CNS). In addition to their ability to produce new cells, NSCs secrete a variety of products, known as the NSC secretome, that have been shown to ameliorate CNS disease pathology and promote recovery. As pre-clinical and clinical research to harness the NSC secretome for therapeutic purposes advances, a more thorough understanding of the endogenous NSC secretome can provide useful insight into the functional capabilities of NSCs. In this review, we focus on research investigating the autocrine and paracrine functions of the endogenous NSC secretome across life. We also compare the NSC secretome across species, finding signs of conserved parallels between rodent, human and zebrafish NSC secretomes. Throughout development and adulthood, we find evidence that the NSC secretome is a critical component of how endogenous NSCs regulate themselves and their niche. We also find gaps in current literature, most notably in the clinically relevant domain of endogenous NSC paracrine function in the injured CNS. Future investigations to further define the endogenous NSC secretome and its role in CNS tissue regulation are necessary to bolster our understanding of NSC-niche interactions and to aid in the generation of safe and effective NSC-based therapies.

Superantigens are unconventional antigens which recognise immune receptors outside the usual binding sites e.g. complementary determining regions (CDRs), to elicit a response within the target cell. T-cell superantigens crosslink T-cell receptors and MHC Class II molecules on antigen-presenting cells, leading to lymphocyte recruitment, induction of cytokine storms and T-cell anergy or apoptosis among many other effects. B-cell superantigens, on the other hand, bind immunoglobulin receptors on B-cells affecting opsonisation, IgG-mediated phagocytosis, and drive B-cells into apoptosis. Here, through a review of the structural basis for recognition of immune receptors by superantigens, we show that their binding interfaces share specific physicochemical characteristics when compared with other protein-protein interaction complexes. Given that antibody-binding superantigens have been exploited extensively in industrial antibody purification, these observations could facilitate further protein engineering to optimize the use of superantigens in this and other areas of biotechnology.

Syncytium formation, i.e., cell-cell fusion resulting in the formation of multinucleated cells, is a hallmark of infection by paramyxoviruses and other important viruses. This natural mechanism has historically been a diagnostic marker for paramyxovirus infection in vivo and is now widely studied for virus-induced membrane fusion in vitro. However, the role of syncytium formation in within-host dissemination and pathogenicity of viruses remains poorly understood. The diversity of henipaviruses and their wide host range and tissue tropism make them particularly appropriate models to characterize the drivers of syncytium formation and its implications for virus fitness and pathogenicity. Based on the henipavirus literature, we summarized current knowledge on the mechanisms driving syncytium formation, mostly acquired from in vitro studies, and on the in vivo distribution of syncytia. While these data suggest that syncytium formation widely occurs across henipaviruses, hosts and tissues, we identified important data gaps that undermined our understanding of the role of syncytium formation in virus pathogenesis. Based on these observations, we propose solutions of varying complexity to fill these data gaps, from better practices in data archiving and publication for in vivo studies, to experimental approaches in vitro.

Circulating atypical cells (CAC) are released from a primary tumour site into peripheral blood and are indicators of cancer metastasis. CAC occur at very low frequency in circulating blood, and their detection remains challenging. Moreover, white blood cells (WBC) are the major contaminant in enriched CAC samples. Here, we developed matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) as a novel CAC characterization platform. Main spectra profiles (MSP) of normal and cancer cells were generated by MALDI-TOF MS, and a cell-main spectra database was then compiled and analysed using the MALDI Biotyper software. Logarithmic scores accurately predicted distinct cell types. The feasibility of this workflow was then validated using simulated samples, which were prepared by 5,000 WBC of three healthy individuals spiked with varying numbers (3, 6, 12, 25, 50 and 100) of lung, colon, or prostate cancer cells. MALDI-TOF MS was able to detect cancer cells down to six cells over the background noise of 5,000 WBC with significantly higher predictive scores as compared to WBC alone. Further development of cell-MSP database to cover all cancer types sourced from cell lines and patient tumours may enable the use of MALDI-TOF MS as a cancer-screening platform in clinical settings in the future.

Cholinergic innervation in pancreas controls both the release of digestive enzymes to support the intestinal digestion and absorption, as well as insulin release to promote nutrient use in the cells of the body. The effects of muscarinic receptor stimulation are described in detail for endocrine beta cells and exocrine acinar cells separately. Here we describe morphological and functional criteria to separate these two cell types in situ in tissue slices and simultaneously measure their response to ACh stimulation on cytosolic Ca2+ oscillations [Ca2+]c in stimulatory glucose conditions. Our results show that both cell types respond to glucose directly in the concentration range compatible with the glucose transporters they express. The physiological ACh concentration increases the frequency of glucose stimulated [Ca2+]c oscillations in both cell types and synchronizes [Ca2+]c oscillations in acinar cells. The pharmacological ACh concentration further increases the oscillation frequency on the level of individual beta cells, inhibits the synchronization between these cells, and abolishes oscillatory activity in acinar cells. We discuss possible mechanisms leading to the observed phenomena.

Purpose: Myelodysplastic syndromes (MDS) are caused by a stem cell failure, but the relationship between immune dysregulation and the course of disease has not yet been analyzed in detail. Experimental design: To get insights into the pathophysiologic and clinical relevance of the histotopography of immune cell subpopulations in this process, the immune cell infiltrate with focus on its spatial distribution was determined by multispectral imaging (MSI) in 147 bone marrow biopsies from MDS or secondary acute myeloid leukemia (sAML) patients and healthy controls (HC). In addition, the data were correlated to genetic alterations and clinical features of these patients including therapy response. Results: A high inter-tumoral heterogeneity in the frequency and spatial distribution of CD3⁺CD8⁺, CD3⁺CD8^-, CD3⁺FOXP3⁺ T cell subsets, MUM1p⁺CD3^- post-germinal B/plasma cells and CD34⁺ blasts was found in MDS and sAML samples. In HC only few B cells/plasma cells, but no T cell subpopulations were detected in the proximity to CD34⁺ blasts. In contrast, the frequency of these lymphocytes was increased in proximity to CD34⁺ blasts in both MDS and sAML independent of the karyotype, genetic alterations frequently detected in MDS, clinical risk stratification systems or treatment response to hypomethylating agents. Furthermore, an increased frequency of CD3⁺CD8⁺ T cells and MUM1p⁺ CD3^- B cells was found in responders to epigenetic drugs. Conclusions: Thus, we conclude that (i) T cell subsets do not belong to the normal stem cell niche, (ii) the presence of T and B cell subpopulations not directly affect the course of MDS, (iii) lymphocytes in the proximity to CD34⁺ blasts might indicate defective stem cell properties and (iv) the number of lymphocytes is a predictor of therapy response to hypomethylating agents.

Cell free systems are a widely used research tool in systems and synthetic biology and a promising platform for manufacturing of proteins and chemicals. In the past, cell free biology was primarily used to better understand fundamental biochemical processes. Notably, E. coli cell free extracts were used in the 1960s to decipher the sequencing of the genetic code. Since then, the transcription and translation capabilities of cell free systems have been repeatedly optimized to improve energy efficiency and protein yield. Today, cell free systems, in combination with the rise of synthetic biology, have taken on a new role as a promising technology for just in time manufacturing of therapeutically important biologics and high-value small molecules. They have also been implemented in an industrial scale for the production of antibodies and cytokines. In this review, we discuss the evolution of cell free systems, advancements in cell free protein synthesis, and cell free metabolic engineering, and conclude with discussing the importance and feasibility of mathematical modeling in cell free systems.

Cell death resistance is a key feature of tumor cells. One of the main anti-cancer therapies is increasing the susceptibility of cells to death. Cancer cells have developed a capability of tumor immune escape. Hence, restoring the immunogenicity of cancer cells can be suggested as an effective approach against cancer. Accumulating evidence proposes that several anticancer agents provoke the release of danger-associated molecular patterns (DAMPs) that are determinants of immunogenicity and stimulate immunogenic cell death (ICD). It has been suggested that ICD inducers are two different types according to their various activities. Here, we review the well-characterized DAMPs and focus on the different types of ICD inducers and recent combination therapies that can augment the immunogenicity of cancer cells.

Angiogenesis, the sprout and growth of new blood vessels from existing vasculature, is an important process of tumor development for the supply of oxygen and nutrition to cancer cells. Endothelial cell is a critical player in angiogenic process by modulating cell proliferation, cell motility, and cell morphology in the response to pro-angiogenic factors and environments provided by tumor and cancer cells. Recent in vivo and in vitro studies have revealed that gap junction of endothelial cells also participates in the promotion of angiogenesis. Pro-angiogenic factors modulate gap junction function and connexins expression in endothelial cells, whereas endothelial connexins involve in angiogenic tube formation and cell migration of endothelial cells via both gap junction channel function dependent or independent mechanisms. In particular, connexin might have the potential to regulate cell mechanics such as cell morphology, cell migration, and cellular stiffness that are dynamically changed during angiogenic processes. Here, we review the implication for endothelial gap junction and cellular mechanics in vascular angiogenesis.

Alterations in gene expressions are often due to epigenetic modifications that can lead to significant influence on cancer development, growth, and progression. The main epigenetic modifications observed in human are methylation and acetylation. In this regard, the HDAC inhibitors (HDACi) such as SAHA (Vorinostat), which can exert epigenetic alterations through impacting the acetylation status of histones, are in clinical trials as a new class of drugs with promising effects on the cancer growth and metastatic process. The small molecule RG7388 is a newly developed inhibitor that is specific for an oncogene-derived protein called MDM2, which is in clinical trials for the treatment of various types of cancers. One of the common characteristics for these two drugs is their ability to induce p21 expression through distinct mechanisms in MCF-7 and LNCaP cells. This difference was expected trigger cell cycle arrest and cell death through intra-cellular mechanisms that are not identical. Hence, the molecular mechanism whereby SAHA can induce cell cycle arrest and trigger necrosis, apoptosis or necroptosis is still evolving. Similarly, the ability of RG7388 for producing anticancer effect is undergoing thorough investigation, since it can produce p53 dependent and p53 independent effects. In this study we performed experiments to measure the cell cycle arrest effects of SAHA and RG7388 on using MCF-7 and LNCaP cells. The cytotoxicity, cell cycle arrest and apoptosis/necroptosis effects of the treatments were assessed by using Trypan Blue Dye Exclusion (TBDE) method, MTT assay, Fluorescence assay with DEVD-amc fluorogenic substrate and Immunoblotting methods. Our results from MCF-7 and LNCaP cells confirmed that SAHA and RG7388 treatments were able to induce cell death via combination of cell cycle arrest and cytotoxic mechanisms. We are speculating that our findings could lead to the development of newer treatments for breast and prostate cancers using this type of combinations.

Enamel matrix derivative (EMD) is used for periodontal tissue regeneration therapy, and can induce mineralization in dental pulp cells (DPCs). We designed a synthetic peptide (SP) derived from the response of cells to EMD, and investigated the effect of the SP on potentiating osteogenesis in DPCs, which have a critical role of dental pulp homeostasis. DPCs were treated with 0, 10, 100, or 1000 ng/mL SP to determine its effect on cell proliferation, cell migration, cell differentiation, and mineralization. We then examined the molecular effects of the SP, focusing on changes in the mitogen-activated protein kinases (MAPK) signaling pathway in these cells. The SP significantly promoted DPC proliferation and migration. Cultures treated with the SP also showed an enhanced expression of markers of osteogenic differentiation and mineralization. The SP also induced the activation of MAPK signaling pathway components. These results suggest that our SP could promote the dental pulp tissue repair by hard tissue formation and the mineralization through activating MAPK signaling pathway. This study provides the first evidence that SP might be a new material for dental pulp tissue treatment.

TTF1-NP(5,2′,4′-trihydroxy-6,7,5′-trimethoxyflavone nanoparticles), derived from the traditional Changbai Mountain medicinal plant Sorbaria sorbifolia (SS), has been showedits anti-cancer effect in various liver cancer cell types and tissues. The present study was designed to evaluate the antitumor mechanism of the TTF1-NP against HepG2 hepatoma cells and HepG2 cells-induced hepatocarcinoma (HCC) in nude mouse model. Here we demonstrated that TTF1-NP inhibits tube formation of HUVECs and HepG2 cell migration and invasion, and inhibits tumor growth in nude mice implanted with HepG2 cells through the downregulation of STAT3 protein and activation, along with VEGF, KDR, bFGF, MMP2 and MMP9 levels. We further revealed that TTF1-NP decreased the DNA-binding capacity of STAT3. Together our results provide a mechanism by which TTF1-NP suppresses cancer cell migration, invasion and angiogenesis through the action of STAT3 and suggests TTF1-NP as a potential therapy for hepatocellular cancer treatment.

NK population is characterized by distinct NK cell subsets which respond differently to the various activating stimuli. For this reason, the determination of the optimal cytotoxic activation of the different NK subsets can be a crucial aspect to be exploited to counter cancer cells in oncologic patients. To evaluate how the triggering of different combination of activating receptors can affect the cytotoxic responses of different NK cell subsets, we developed a microbead-based degranulation assay. By using this new assay, we were able to detect CD107a+ degranulating NK cells even within the less cytotoxic subsets (i.e. resting CD56bright and unlicensed CD56dim NK cells), thus demonstrating its high sensitivity. Interestingly, signals delivered by the co-engagement of NKp46 with 2B4, but not with CD2 or DNAM-1, strongly cooperate to enhance degranulation on both licensed and unlicensed CD56dim NK cells. Of note, 2B4 is known to bind CD48 hematopoietic antigen, therefore this observation may provide the rationale why CD56dim subset expansion correlates with successful hematopoietic stem cell transplantation mediated by alloreactive NK cells against host T, DC and leukemic cells, while sparing host non-hematopoietic tissues and graft versus host disease. The assay further confirms that activation of LFA-1 on NK cells leads to their granule polarization, even if, in some cases, this also takes to an inhibition of NK degranulation, suggesting that LFA-1 engagement by ICAMs on target cells may differently affect NK cell response. Finally, we observed that NK cells undergo a time-dependent spontaneous (cytokine-independent) activation after blood withdrawal, an aspect that may strongly bias the evaluation of the resting NK cell response. Altogether our data may pave the way to develop new NK activation and expansion strategies that target the highly cytotoxic CD56dim NK cells and can be feasible and useful for cancer and viral infection treatment.

Products made from petroleum-derived plastic materials are linked to many environmental problems such as greenhouse gas emissions and plastic pollution. It is desirable to manufacture products from environmentally friendly materials instead of petroleum-based plastic materials. Products made from biomass-fungi composite materials are biodegradable and can be utilized for packaging, construction, and furniture. In biomass-fungi composite materials, biomass particles (derived from agricultural wastes) serve as the substrate and the fungal hyphae network binds the biomass particles together. There are many reported studies on 3D printing of biomass-fungi composite materials. However, there are no reported studies on biodegradation of 3D printed samples from biomass-fungi composite materials. In this study, two types of biomass materials were used to prepare printable mixture hemp hurd and beechwood sawdust. The fungi strain used was Trametes versicolor. Extrusion based 3D printing was used to print samples. 3D printed samples were left for five days to allow fungi to grow. The samples were then dried in an oven for 4 hours at 120ºC to kill all the fungi in the samples. The samples were buried in the soil using a mesh bag and kept in an environmental chamber at 25℃ with a relative humidity of 48%. The weight of these samples was measured every week over the period of three months. Both types of samples (prepared from beechwood sawdust and hemp hurd, respectively) showed weight change after three months. Furthermore, the samples made from hemp hurd had more weight change than the samples made from beechwood sawdust. The SEM (scanning electron microscope) micrographs and FTIR (Fourier transform infrared) spectroscopy showed evidence of biodegradation of these samples.

Background: In high-risk breast cancer patients with skin infiltration the administration of a uniform dose to superficial tissues is fundamental in order to reduce local skin relapse. A personalized bolus may prevent the potential standard bolus’ inadequate dose distribution due to air gaps between the bolus and the skin. In this pilot study, we introduced in clinical practice the use of a personalized 3D printed bolus filled with ultrasound transmission gel. Methods: Seven patients undergoing radiotherapy after mastectomy were selected. 3D printed bolus dosimetric assessment was performed with MOSFET dosimeters on an anthropomorphic phantom and subsequently on three selected cases with increasing bolus shape irregularity. Acute/late toxicity and local control was assessed. Results: Overall, for the clinical cases the percentage median difference between measured and calculated doses was -2.7% (-7.0%÷4.9%). Median follow-up was 21 months. After two years, one patient showed G2 pain, one patient manifested G1 telangiectasia, one patient showed G1 hyperpigmentation, and two patients had no relevant toxicity. Conclusions: Personalized 3D-printed bolus filled with ultrasound gel may easily reproduce the standard bolus’ consistency providing an accurate coverage of the target area with tolerable acute/late toxicity grades. This is a pilot study and further investigations are needed.

On using a 3D model to teach students a topic in astronomy, feedback was evaluated via a questionnaire. Analysis of students' responses to the questions revealed that user interface had a significant impact on their attitudes, which in turn could increase users' willingness to use immersive technology in e-learning. The results also showed that improving visual features through immersive technology and augmented reality tools were effective. The results confirm all hypotheses about positive relationships between facilitating conditions and usability of metaverse technology for students' attention and study motivation.

Drone technology has undoubtedly become an integral part of our everyday life these days. The business and industrial use of unmanned aerial vehicles (UAV) can provide advantageous solutions in many areas of life, and they are also optimal for emergency situations and for accessing hard-to-reach places. However, their application poses numerous technological and regulatory challenges to be overcome. One of the weak links in the operation of UAVs is the limited availability of energy. In order to address this issue authors developed a novel trajectory planning method for UAVs to optimize the energy necessity during flight. First, an “energy map” was created, which was the basis for trajectory planning, i.e., determining the energy consumption of the individual components. This was followed by configuring the 3D environment including partitioning of the work space (WS), in fact, defining the free spaces, occupied spaces (obstacles) and semi-occupied/free spaces. Then the corresponding graph-like path(s) were generated on the basis of the partitioned space, where a graph search-based heuristic trajectory planning was initiated, taking into account the most important wind conditions including velocity and direction. Finally, in order to test the theoretical results, some sample environments were created to test and analyse the proposed path generations. The method eventually proposed was able to determine the optimal path in terms of energy consumption.

This study aimed to compare the flexural strength, surface roughness, and microhardness of a resin for three-dimensional (3D) printing and a conventional composite resin and to evaluate whether the printing orientation influences these properties. To evaluate the flexural resistance, test specimens were produced and divided into four groups: three groups of resins for 3D printing with inclinations of 0°, 45°, and 90°, and one group of conventional composite resin. Forty discs were produced and subjected to a sandpaper-polishing sequence, and the surface roughness was measured using a roughness meter. The Vickers microhardness test was performed at three different points, and the average was obtained. The results were subjected to ANOVA statistical analysis and Tukey's test. There were statistical differences in the flexural strength and microhardness between the conventional resin and the resin used for 3D printing. No statistical difference in surface roughness was observed. The resin for 3D printing exhibited lower flexural strength and microhardness than conventional composite resins. We concluded that the resin for 3D printing had lower flexural strength and hardness than the conventional composite resin but similar surface roughness. The printing orientation did not affect the flexural strength, whereas the hardness increased as the printing angle increased.

Background and Objectives To compare navigator-gated three-dimensional T1-weighted gradient-echo sequences (NG-3D T1W-GRE) with standard breath-hold T1W-GRE (BH T1W-GRE) for the detection of HCC nodules in patients with chronic viral hepatitis which undergo at magnetic resonance imaging (MRI) using gadoxetic acid. Methods and materials We evaluated 58 patients with hepatic nodules at least 1 cm in diameter detected by ultrasound who underwent liver MRI including NG-3D T1W-GRE sequences and breath hold T1-weighted gradient-echo sequences with different flip angles(FA) at 10-25-40° after contrast administration of gadoxetic acid at magnetic scanner operating at1.5 Tesla. Image quality and relative hepatic and nodular enhancement were evaluated by three blinded reviewers. Results In these patients, image quality of NG-3D T1W-GRE was assessed significantly better than quality of BHT1W- GRE (P < 0.005) and relative enhancement was significantly higher than that of BH- T1W-GRE ( P

Fabrication of scaffolds via 3D printing is a promising approach for tissue engineering. In this study, we combined 3D printing with cryogenic crosslinking to create biocompatible gela-tin/oxidized alginate (Gel/OxAlg) scaffolds with large pore sizes, beneficial for bone tissue re-generation. To enhance the osteogenic effects and mechanical properties of these scaffolds, we evaluated the impact of hydroxyapatite (HAp) on the rheological characteristics of the 2.86% (1:1) Gel/OxAlg ink. We investigated the morphological and mechanical properties of scaffolds with low, 5% and high 10% HAp content, as well as the resulting bio and osteogenic effects. Scanning electron microscopy revealed a reduction in pore sizes from 160-180 µm (HAp-free) to 120-140 µm for both HAp-containing scaffolds. Increased stability and higher Young’s moduli were measured for 5% and 10% HAp (18 and 21 kPa, respectively) compared to 11 kPa for HAp-free constructs. Biological assessments with mesenchymal stem cells indicated excellent cytocompati-bility and osteogenic differentiation in all scaffolds, with high degree of mineralization in HAp-containing constructs. Scaffolds with 5% HAp exhibit improved mechanical characteristics and shape fidelity, demonstrate positive osteogenic impact, and enhance bone tissue formation. Increasing the HAp content to 10% did not show any advantages in osteogenesis, offering a minor increase in mechanical strength at the cost of significantly compromised shape fidelity.

The aim of this research is to investigate the influence of model height employed in the deep drawing of orthodontic aligner sheets on force transmission and aligner thickness. Forty sheets (Zendura FLX) were thermoformed over four models of varying heights (15, 20, 25, and 30 mm). Normal contact force generated on the facial surface of the upper right central incisor (Tooth 11) was measured using pressure-sensitive films. Aligner thickness around Tooth 11 was measured at five points. A digital caliper and a micro-computed tomography (µ-CT) were employed for thickness measurements. The normal contact force exhibited an uneven distribution across the facial surface of Tooth 11. Model 15 displayed the highest force (88.9±23.2 N), while Model 30 exhibited the lowest (45.7±15.8 N). The force distribution was more favorable for bodily movement with Model 15. Thickness measurements, conducted using a digital caliper and µ-CT, revealed substantial thinning of the aligner after thermoforming. This thinning was most pronounced at the incisal edge (50% of the original thickness) and least at the gingivo-facial part (85%). Additionally, there was a progressive reduction in aligner thickness with increasing model height, which was most significant on the facial tooth surfaces. We conclude that the thermoplastic aligner sheets undergo substantial thinning during the thermoforming process, which becomes more pronounced as the height of the model increases. As a result, there is a decrease in both overall and localized force transmission, which could lead to increased tipping by the aligner and a diminished ability to achieve bodily movement.

Fingerprint recognition systems have achieved widespread integration into various technological devices, including cell phones, computers, door locks, and time attendance machines. Nevertheless, individuals with worn fingerprints encounter challenges when attempting to unlock original fingerprint systems, resulting in disruptions to their daily activities. This study explores two distinct methods for fingerprint backup: traditional fingerprint impression and 3D printing technologies. Unlocking tests were conducted on commonly available optical fingerprint lock-equipped cell phones to assess the efficacy of these methods, particularly in unlocking worn fingerprints. The research findings indicated that traditional fingerprint impression method exhibited high fidelity in reproducing fingerprint patterns, achieving an impressive unlocking success rate of 97.8% for imprinting unworn fingerprints. However, when dealing with worn fingerprints, traditional fingerprint impression technique showed a reduced unlocking success rate, progressively decreasing with increasing degrees of finger wear. In contrast, 3D printed backup fingerprints, with image processing and optimization of ridge height, mitigated the impact of fingerprint wear on unlocking capability, resulting in an unlocking success rate of 84.4% or higher. Thus, the utilization of 3D printing technology proves advantageous for individuals with severely worn or incomplete fingerprints, providing a viable solution for unforeseen circumstances.

The assortment of low-cost 3D printers for “in-practice” use, e.g., for clear aligner therapies, is ever increasing. To address concerns about the accuracy of orthodontic models produced on such printers when cost efficient modes of 3D printing are employed, this study examined the effect of print model body type and print angulation on accuracy. Six printing-configuration groups were included: 2 model types (solid or hollow shell) combined with 3 print angles (0°, 70°, or 90°) with 10 models/group; all models were printed with 100-µm layer thickness using a digital light processing-based 3-dimensional printer. Eleven selected structures and distances were measured on the printed models with a digital microscope and compared to the same measures on a digitized master model. The clinically-acceptability range was set at ±0.25 mm difference from the master model for single tooth measurements (intra-tooth) and ±0.5 mm for cross-arch measurements (inter-tooth). For individual measurements across all models, 98% fell within clinical acceptability. For mean measurements within each model group, only canine height for the shell-0° model had a mean difference (0.26 mm ± 0.03) outside the clinically acceptable range for intra-tooth measurements. Standard deviations for all intra-tooth measurements were within 0.07 mm. While none of the mean inter-tooth measurements exceeded the acceptability range, the standard deviations were larger (0.04 to 0.30 mm). Accuracy of orthodontic models for clear aligner therapies was not impacted beyond the clinically acceptable range when altering model body type and print angulation to improve efficiency of 3D printing. These findings suggest greater flexibility of the practitioner to alter print settings to address time and cost efficiency in various clinical scenarios and still maintain clinically acceptable model accuracy.

With the development of organizational biology and forced from the EU Cosmetic Regulation (EU/1223/2009) with the complete ban of animal testing for cosmetic purposes, 3D skin models have been widely developed due to their similar physiological structure and metabolic function to native human tissue. 3D skin models for in vitro cosmetic evaluation are still promising alternative methods for evaluating phototoxicity, corrosivity and irritation. These models have been used for early screening and later validation of toxicity and efficacy evaluation by many well-known cosmetic companies. 3D skin models have become an important tool in cosmetic evaluation research. This article reviews the development of 3D skin models and their application in cosmetic evaluation. An outlook on the current application and future technological development directions in the cosmetic field is also presented.

Implementing digital technologies, such augmented reality and 3D models, to various cultural sectors is an ongoing procedure, especially favorable after COVID-19 pandemic. Even though such technologies are familiar to museums, galleries and archaeological places all over the world, little research or practical cases have been developed when it comes to theaters and theatrical costumes. Proposed research article describes the methodology of both reference material collection and applications development, while an analysis of functions and aesthetic results are described, especially when it comes to 3D models. The paper’s aim to reveal that theatrical operations (as part of cultural sector) as well as structured parts (such as acts, performances and costumes) can be preserved and served to larger audiences via technological means, such as an augmented reality application.

This pioneering study presents an in-depth biomechanical examination of soccer's diving header, aiming to quantify its impact on ball speed enhancement (BSE) and effective offensive range (EOR). Despite the diving header's widespread acclaim and historical significance, there remains a dearth of scientific scrutiny into its biomechanical intricacies. Employing an innovative research design featuring a static hanging ball at varied offensive distances and heights, the study replicates diverse header scenarios. The results of 3D motion quantification have shown that a physically excellent player (identified through the maximal standing long jump test) could reach an EOR around 2.64 times his body height. Furthermore, the study unveils that proficient players could attain BSE surpassing 9 m/s, signifying the diving header's heightened efficacy compared to traditional heading techniques, which could only result in 4.5 m/s. Correlation analyses unveil noteworthy relationships, highlighting the pivotal role of head speed at impact and the influence of minimizing speed-drop and temporal disparities for amplified effectiveness. Considerations for optimizing diving header execution are introduced, emphasizing the necessity for targeted training programs. Despite acknowledged limitations inherent to its pilot nature, this exploration furnishes foundational knowledge to guide subsequent research and practical applications, providing valuable insights into soccer training and skill development through a biomechanical lens.

This paper introduces an innovative approach to lipreading, leveraging a web application designed to generate subtitles for videos where the speaker's mouth is visible and a comprehensive literature review that precedes the discussion, encompassing a thorough examination of various lipreading methods employed over the past decade. Our method employs a powerful deep learning model, featuring a 3D-convolution network and bidirectional LSTM, enabling accurate sentence-level predictions based solely on visual lip movements. With an impressive accuracy of 97%, our model is trained using pre-segmented lips regions, transformed into animated GIFs for effective pre-training. This work stands as a significant contribution to the evolving landscape of lipreading research, offering a practical and accurate solution for real-world applications.

Background: Anthropometric measurements play a crucial role in medico-legal practices. Actually, several scanning technologies are employed in post-mortem investigations for forensic anthropological measurements. This study aims to evaluate the precision, in-ter-rater reliability, and accuracy of a hand-held scanner in measuring various body parts. Methods: Three independent raters measured seven longitudinal distances using an iPad Pro equipped with a LiDAR sensor and specific software. These measurements were sta-tistically compared to manual measurements conducted by an operator using a laser level and a meterstick (considered the gold standard). Results: The Friedman’s test revealed minimal intra-rater variability in digital measurements. Inter-rater variability analysis yielded an ICC=1, signifying high agreement among the three independent raters. Addi-tionally, the accuracy of digital measurements displayed errors below 2%. Conclusions: Preliminary findings demonstrate that the pairing of LiDAR technology with the Polycam app showcases high precision, inter-rater agreement, and accuracy. Hand-held scanners show potential in forensic anthropology due to their simplicity, affordability, and porta-bility. However, further validation studies under real-world conditions are essential to es-tablish the reliability and effectiveness of handheld scanners in medico-legal settings.

Real-time multi-axis distributed tactile sensing is a critical capability if robots are to perform stable gripping and dexterous manipulation, as it provides crucial information about the sensor-object interface. In this paper, we present an optical-based 6-axis tactile sensor designed in a fingertip shape for robotic dexterous manipulation. The distributed sensor can precisely estimate local XYZ force and displacement at ten distinct locations, and provide global XYZ force and torque measurements. Its compact size, comparable to that of a human thumb, and minimal thickness allow seamless integration onto existing robotic fingers, eliminating the need for complex modifications to the gripper. The proposed sensor design uses a simple, low-cost fabrication method. Moreover, the optical transduction approach uses light angle and intensity sensing to infer force and displacement from deformations of individual sensing units that form the overall sensor, providing distributed 6-axis sensing. The local force precision at each sensing unit in the X, Y, and Z axes is 20.89 mN, 19.19 mN, and 43.22 mN, respectively, over an local force range of approximately ±1.5 N in X and Y and 0 to -2 N in Z. The local displacement precision in the X, Y and Z axes is 56.70 μm, 50.18 μm and 13.83 μm, respectively, over a local displacement range of ±2 mm in the XY directions and 0 to -1.5mm in Z (i.e., compression). Additionally, the sensor can measure global torques, Tx, Ty and Tz, with a precision of of 1.90 N-mm, 1.54 N-mm and 1.26 N-mm, respectively. The fabricated design is showcased by integrating it with an OnRobot RG2 gripper and illustrating real-time measurements during in simple demonstration task which generate changing global forces and torques.

Bacteriotherapy is emerging as a strategic and effective approach to treat infections by providing putatively harmless bacteria (i.e., probiotics) as antagonists to pathogens. A proper delivery of probiotics or their metabolites (i.e., post-biotics) can avail itself of biomaterial encapsulation by innovative manufacturing technologies. This review paper aims at providing the most recent biomaterial-assisted strategies proposed to treat infections or disbiosis using bacteriotherapy. We revised the encapsulation processes across multiscale biomaterial approaches, which could be ideal to target different tissues and suit diverse therapeutic opportunities. Hydrogels, and in particular polysaccharides, are the focus of this review, as they have been reported to better sustain vitality of the live cells incorporated. Specifically, the approaches for fabricating hydrogel-based devices with increasing dimensionality (D), namely, 0D (i.e., particles), 1D (i.e., fibers), 2D (i.e., fiber meshes), and 3D (i.e., scaffolds) endowed with probiotics, were detailed by describing their advantages and challenges, along with a future overlook in the field. Electrospinning, electrospray and 3D bioprinting were investigated as new biofabrication methods for probiotic encapsulation within multidimensional matrices. Finally, examples of biomaterial-based systems for cell and post-biotic release were reported.

Accurate weight measurement is pivotal for monitoring the growth and well-being of cattle. However, the conventional weighing process, which involves physically placing cattle on scales, is labor-intensive and distressing for the animals. Hence, the development of automated cattle weight prediction techniques assumes critical significance. This study proposes a weight prediction approach for Korean cattle using 3D segmentation-based feature extraction and regression machine learning techniques from incomplete 3D shapes acquired from real farm environments. In the initial phase, we generated mesh data of 3D Korean cattle shapes using a multiple-camera system. Subsequently, deep learning-based 3D segmentation with the PointNet network model was employed to segment two dominant parts of the cattle. From these segmented parts, three crucial dimensions of Korean cattle were extracted. Finally, we implemented five regression machine learning models (CatBoost regression, LightGBM, Polynomial regression, Random Forest regression, and XGBoost regression) for weight prediction. To validate our approach, we captured 270 Korean cattle in various poses, totaling 1190 poses of 270 cattle. The best result was achieved with mean absolute error (MAE) of 25.2 kg and mean absolute percent error (MAPE) of 5.81% using the random forest regression model.

Type 1 diabetes is a chronic condition that results from the destruction of insulin-producing β-cells in the pancreas. Current treatments for type 1 diabetes, such as insulin therapy and pancreatic islet transplantation, have several limitations and, hence not quite effective in the long run. As current therapy methods fail to slow disease development, novel strategies such as the development of a bioartificial pancreas are being seriously considered. Over the last decade, research has focused on tissue engineering, which aids in the design of biological alternatives for the repair and replacement of non-functional or damaged organs. Three dimensional (3D) bioprinting technology which employs 3D printing technology to generate 3D tissue-like structures from biomaterials and cells, offers a promising solution for the treatment of type 1 diabetes by providing the ability to generate functional endocrine pancreatic tissue. Bioprinted structures are therefore an important aspect of tissue engineering because they have been found to replicate the native extracellular matrix, promoting cell survival and proliferation. In this review, we extensively discussed about bioprinting endocrine pancreas for the treatment of type 1 diabetes particularly focussing on the choice of cells, biomaterials, growth factors, and essential considerations. Additionally, the key challenges and perspectives towards recapitulation of the pancreatic function of the pancreatic organ engineering technologies have also been discussed.

Coronary artery disease is the most prevalent cardiovascular disease, claiming millions of lives annually around the world. The current treatment includes surgically inserting a tubular construct called as a stent inside arteries to restore blood flow. However, due to lack of patient-specific design, the commercial products cannot be used with different vessel anatomies. In this review, we have summarized the drawbacks in existing commercial metal stents which face problems of restenosis and inflammatory responses, owing to the development of neointimal hyperplasia. Further, we have highlighted the fabrication of stents using biodegradable polymers, which can circumvent most of the existing limitations. In this regard, we elaborated on utilization of new fabrication methodologies based on additive manufacturing such as three-dimensional printing to design patient-specific stents. Finally, we have discussed on the functionalization of these stent surfaces with suitable bioactive molecules which can prove to enhance their properties in preventing thrombosis and better healing of injured blood vessel lining.

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.

This paper proposes a multimodal fusion 3D target detection algorithm based on the attention mechanism to improve the performance of 3D target detection. The algorithm utilizes point cloud data and information from camera. For image feature extraction, the ResNet50+FPN architecture extracts features at four levels. Point cloud feature extraction employs the voxel method and FCN to extract point and voxel features. The fusion of image and point cloud features is achieved through regional point fusion and regional voxel fusion methods. After information fusion, the Coordinate attention mechanism and SimAM attention mechanism extract fusion features at a deep level. The algorithm's performance is evaluated using the DAIR-V2X dataset. The results show that compared to the Part-A2 algorithm, the proposed algorithm improves the mAP value by 7.9% in BEV view and 7.8% in 3D view at IOU=0.5 (cars) and IOU=0.25 (pedestrians and cyclist). At IOU=0.7 (cars) and IOU=0.5 (pedestrians and cyclist), the mAP value of the SECOND algorithm is improved by 5.4% in the BEV view and 4.3% in the 3D view, compared to other comparison algorithms.

3D/4D printing technologies are currently among the fastest growing, cutting edge fabrication technologies. The scale of their applications is vast and applicable to nearly all industries. 3D printing technologies are particularly popular in robotics, and especially in advanced design innovative solutions for areas such as manufacturing, space technology and medicine. The development of robotics, and in particular of the precision of manufactured components, such as actuators, pneumatic muscles, power transmission units, etc., means that new prototypes are still being made, where the use of 3D printers reduces the production time severalfold and allows for completing the necessary simulations and tests. In addition, the use of 3D printers allows for the production of thin-walled and cellular structures, which is a great advantage compared to conventional fabrication technologies. In the range of 3D printers available on the market, only a few selected technologies allow for actual use in the construction of advanced robot elements (muscles, vibration dampers, etc.). In the era of rapid growth of the precision of the available 3D printers and modern materials, 3D printing may soon become a major tool in robotics. This article presents an overview of 3D printing technologies and materials in terms of their application in robotics and provides examples of the use of 3D and 4D printing in prototyping and fabrication of robotic elements with particular emphasis on the current state of the art. The study considered the possibilities of using 3D/4D printing in robotics with the use of polymeric materials. The review of the literature and the research work currently being carried out in this area is very promising and it seems that 3D/4D printing in robotics is widely used and is still developing, which allows to conclude that in the near future the number of research works in this field will increase rapidly.

Selective laser sintering (SLS) has drawn attention for the fabrication of three-dimensional oral dosage forms due to the plurality of drug formulations that can be processed. The aim of this work was to employ SLS with a CO2 laser for the manufacturing of carvedilol personalised dos-age forms of various strengths. Carvedilol (CVD) and vinylpyrrolidone-vinyl acetate copolymer (Kollidon VA64) blends of various ratios were sintered to produce CVD tablets of 3.125, 6.25 and 12.5 mg. The tuning of the SLS processing parameter laser intensity, improved printability and impacted the tablet hardness, friability, CVD dissolution rate, and the total amount of drug re-leased. Physicochemical characterization showed the presence of CVD in the amorphous state. X-ray micro-CT analysis demonstrated that the applied CO2 intensity affected the total tablet po-rosity, which was reduced with increased laser intensity. The study demonstrated that SLS is a suitable technology for the development of personalised medicines that meet the required specifications and patient needs.

Two antimicrobial agents such as silver nanoparticles (AgNPs) and titanium dioxide (TiO2) have been formulated with natural polysaccharides (chitosan or alginate) to develop innovative inks for the rapid, customizable, and extremely accurate manufacturing of 3D printed scaffolds useful as dressings in the treatment of infected skin wounds. Suitable chemical-physical properties for the applicability of these innovative devices were demonstrated through the evaluation of water content (88-93%), mechanical strength (Young’s modulus 0.23-0.6 MPa), elasticity, and morphology. The antimicrobial tests performed against Staphylococcus Aureus and Pseudomonas Aeruginosa demonstrated the antimicrobial activities against Gram+ and Gram– bacteria of AgNPs and TiO2 agents embedded in the CH or ALG macroporous 3D hydrogels (AgNPs MIC starting from 5 µg/ml). The biocompatibility of chitosan was widely demonstrated by cell viability tests and was higher than that observed for alginate. Constructs containing AgNPs at 10 µg/ml concentration level did not significantly alter cell viability as well as the presence of titanium dioxide; cytotoxicity towards human fibroblasts was observed starting with AgNPs concentration of 100 µg/ml. In conclusions, the 3D printed dressings here developed own the features to be cheap, highly defined, easy to be manufactured and further applied in personalized antimicrobial medicine applications.

3D bioprinting is the process of manipulating cell-laden bioinks to fabricate living structures. 3D bioprinting techniques have brought considerable innovation in biomedicine, especially in the field of tissue engineering, allowing the production of 3D organ and tissue models for in vivo transplantation purposes or for in-depth and precise in vitro analyses. Natural-derived hydrogels, especially those obtained from the decellularization of biological tissues, are promising bioinks for 3D printing purposes, as they present the best biocompatibility characteristics. Despite this, many natural hydrogels do not possess the necessary mechanical properties to allow a simple and immediate application in the 3D printing process. In this review, we focus on the bioactive and mechanical characteristics that natural hydrogels may possess to allow efficient production of organs and tissues for biomedical applications, emphasizing the reinforcement techniques to improve their biomechanical properties.

Recently, new semantic segmentation and object detection methods have been proposed for direct processing of three-dimensional (3D) LiDAR sensor point clouds. LiDAR can produce highly accurate and detailed 3D maps of natural and man-made environments and is used for sensing in many contexts due to its ability to capture more information and its robustness to dynamic changes in the environment. In addition, the cost of LiDAR sensors has decreased in recent years, which is an important factor for many application scenarios. The challenge with 3D LiDAR sensors like the Velodyne HDL-64E S2 is that they can output large 3D data at up to 1,300,000 points per second, which is difficult to process in real time when applying complex algorithms and models for efficient semantic segmentation. Most existing approaches are either only suitable for relatively small point clouds or rely on computationally intensive sampling techniques to reduce their size. As a result, most of these methods do not work in real-time in realistic field robotics application scenarios, making them unsuitable for practical applications. Systematic point selection is a possible solution to reduce the amount of data to be processed, but although it is memory and computationally efficient, it selects only a small subset of points, which may result in important features being missed. To address this problem, we propose a new approach to semantic segmentation in forestry that uses a systematic sampling method in which the local neighbors of each point are retained to preserve geometric details. Our approach has been shown to process up to 1 million points in a single pass. It outperforms the state of the art in efficient semantic segmentation in large datasets such as Semantic3D. We also present a preliminary study on the performance of LiDAR-only data, i.e., intensity values from LiDAR sensors without RGB values for semi-autonomous robot perception.

Corneal opacity is a leading cause of vision impairment and suffering worldwide. Transplantation can effectively restore vision and reduce chronic discomfort. However, there is a considerable shortage of viable corneal graft tissues. Tissue engineering may address this issue by advancing xeno- keratoplasty as a viable alternative to conventional keratoplasty. In particular, livestock decellularization strategies offer the potential to generate bioartificial ocular prosthetics in sufficient supply to match existing and projected needs. To this end, we have examined the best practices and characterizations that have supported the current state-of-the-art driving preclinical and clinical applications. Identifying the challenges that delimit activities to supplement the donor corneal pool derived from acellular scaffolds allowed us to hypothesize a model for keratoprosthesis applications derived from livestock combining 3D printing and decellularization.

This paper introduces a novel 3D NoC router that combines buffered and bufferless routing with approximate priority comparison when deflecting flits. Our proposal is a modification of an asymmetrical router that is buffered in the z dimension ports and bufferless in the x and y dimension ports. Flits that request output ports in the x and y dimensions are granted or deflected based on approximate, instead of accurate, priority comparison. Experimental results show that the proposed router, besides effectively combining the advantages of both buffered and bufferless routers, it achieves additional performance and area gains due to the reduced logic required for approximate priority comparison in flit deflections. Experimental results using synthetic and realistic traffic show that the proposed router begins to saturate at a sifnificantly higher injection rate than a bufferless router, but at a slightly lower injection rate than when using accurate priority comparison. Furthermore, the proposed router achieves higher clock frequencies and reduced area compared to the simpler permutation network.

There has been considerable research on reconstructing 3D shapes from single-view images; however, preserving the detailed information of the input image remains difficult. In this paper, we propose the application of a gradient map to train a network, aimed at improving the visual quality of fine-grained details such as the thin and tiny components of generated shapes. Each gradient map was created from the original voxel data, and each value represented the amount of information per volume. Here, the gradient map was defined by several methods that mathematically quantify and represent the detailed structure of an object. By applying this map to the loss function in training, we could induce the network to intensively train partial details, such as thin and narrow parts. We demonstrated that the detailed information was well-recovered when a weight that is proportional to the gradient value was applied to the loss. Furthermore, it is expected that our method will contribute to the development of 3D technologies related to the construction of virtual space for simulation and new customer experience.

Additive manufacturing or 3D printing applying polycaprolactone-(PCL)-based medical devices represents an important branch of tissue engineering, where the sterilization method is a key process for further safe application in vitro and in vivo. In this study, the authors intend to access the most suitable gamma radiation conditions to sterilize PCL-based scaffolds in a preliminary biocompatibility assessment, envisioning future studies for airway obstruction conditions. Three radiation levels were considered, 25 kGy, 35 kGy and 45 kGy and evaluated as to their cyto- and biocompatibility. All three groups presented biocompatible properties, indicating an adequate sterility condition. As for the cytocompatibility analysis, devices sterilized by 35 kGy and 45 kGy showed better results, with the 45 kGy showing overall improved outcomes. This study allowed to select the most suitable sterilization condition for PCL-based scaffolds, aiming at immediate future assays, by applying 3D-customized printing techniques to specific airway obstruction lesions of the trachea.

This retrospective study evaluated changes in the pharyngeal portion of the upper airway in pa-tients with constricted and normal airway treated with clear aligners (Invisalign, Align). Additionally, the paper has assessed the change of tongue position in the oral cavity from lateral view. Evaluation was performed with specialized software (Invivo 6.0, Anatomage) on pre-treatment and posttreatment pairs of cone beam computed tomography imaging (CBCT) data. The level of airway constriction, volume, cross-section minimal area, and tongue profile were evaluated. Patients with malocclusion, with pair or initial and finishing CBCT and without sig-nificant weight change between the scans, treated with Invisalign clear aligners were distributed in two groups. Group A consisted of fifty-five patients with orthodontic malocclusion and con-stricted upper airway. Control group B consisted of thirty-one patients with orthodontic malocclusions without any airway constriction. In the group with airway constriction, there was a statistically significant increase in volume during therapy (p<0.001). The surface of the most con-stricted cross-section of airway did not change significantly after treatment in any of the groups. The airway constriction was most frequently localized at the level of 2nd cervical vertebra. The final tongue position was different from initial in 62.2% of all clear aligner treatments.

In engineering, connections between components are often weak areas. Unreasonable connection methods can easily reduce the strength of components, resulting in unpredictable failure modes. In nature, numerous connection methods for biological structures with excellent mechanical properties have evolved. Studying the connection methods of organisms in nature can inspire new ideas for bionic connection methods. When the diabolical ironclad beetle is under pressure, the elytra are not easy to separate, which ensures the stability of the beetle's external structure, thus making the beetle extremely resistant to pressure. The reason for this is the interlocking and toughening effect of the unique jigsaw connection between the elytra. Therefore, in this paper a theoretical analysis model is established and used to analyze the mechanical behavior of the diabolical ironclad beetle's jigsaw connection during the drawing process and determine the influence of factors such as quantity, angle, and geometric characteristics on the mechanical properties of the jigsaw connection. The results of the theoretical analysis are then compared with the results of experiments and ABAQUS finite element simulation.

In surveying engineering tasks, close-range photogrammetry belongs to leading technology considering different aspects like the achievable accuracy, availability of hardware and software, accessibility to measured objects, or the economy. Hence, constant studies on photogrammetric data processing are desirable. Especially in industrial applications, the control points for close-range photogrammetry are usually measured using total stations. In the case of small objects, more precise positions of control points can be obtained by deploying and adjusting a three-dimensional linear network set up on the object. The article analyzes the accuracy of the proposed method, based on the measurement of the linear network using a tape with a precision of ±1 mm. The experiment shows that the adjusted positions of the network control points can be determined with high, one-millimeter accuracy. The photogrammetric 3D model derived referring to such control points and stereo-images captured with a non-metric camera is also characterized by the highest possible precision, which qualifies the presented method to accurate measurements used in surveying engineering. The authors prove that the distance between two randomly optional points derived from the 3D model of a dimensioned object is equal to the actual distance measured directly on it with one-millimeter accuracy.

Polymer adhesives have emerged as a promising dielectric passivation layer in hybrid bonding for 3D integration while they raise misalignment problems during curing. In this work, the synergistic effect of oxygen plasma surface activation and wetting is utilized to achieve bonding between completed cured polyimides. The optimized process achieves a void-less bonding with a maximum shear strength of 35.3 MPa at a low temperature of 250 °C in merely 2 min, significantly shortening the bonding period and decreasing thermal stress. It is found that the plasma activation generated hydrophilic groups on the polyimide surface, and the wetting process further introduced more -OH groups and water molecular on the activated polyimide surface. The synergistic process of plasma activation and wetting facilitate bridging polyimide interfaces to achieve bonding, providing an alternative path for adhesive bonding in 3D integration.

Desktop fused filament fabrication (FFF) 3D printers have been growing in popularity among hobbyist and professional users as a prototyping and low-volume manufacturing tool. One issue these printers face is the inability to determine when a defect has occurred rendering the print unusable. Several techniques have been proposed to detect such defects but many of these approaches are tailored to one specific fault (e.g., filament runout/jam), use expensive hardware such as laser distance sensors, and/or use machine vision algorithms which are sensitive to ambient conditions, and hence can be unreliable. This paper proposes a versatile, reliable, and low-cost system, named MTouch, to detect millimeter-scale defects that tend to make prints unusable. At the core of MTouch is an actuated contact probe designed using a low-power solenoid, magnet, and hall effect sensor. This sensor is used to check for the presence, or absence, of the printed object at specific locations. The MTouch probe demonstrated 100% reliability, which was significantly higher than the 74% reliability achieved using a commercially available contact probe (the BLTouch). Additionally, an algorithm was developed to automatically detect common print failures such as layer shifting, bed separation, and filament runout using the MTouch probe. The algorithm was implemented on a Raspberry Pi mini-computer via an Octoprint plug-in. In head-to-head testing against a commercially available print defect detection system (The Spaghetti Detective), the MTouch was able to detect faults 44% faster on average while only increasing the print time by 8.49%. In addition, MTouch was able to detect faults The Spaghetti Detective was unable to identify such as layer shifting and filament runout/jam.

This study presents the results of a research project financed by the Lazio Regional Government. The research focused on defining an integrated model of recent alluvial deposits in the Tiber River. To achieve this objective, geological boreholes were made to monitor the aquifer and in situ and laboratory tests carried out. The data obtained was used to detail stratigraphic aspects and improve the comprehension of water circulation beneath the recent alluvial deposits of the Tiber River in the urban area of Rome, between the Ponte Milvio bridge and the Tiber Island. The stratigraphic intervals recognised in the boreholes were parameterised based on their litho-technical characteristics. The new data acquired, and integrated with existing data in the CNR IGAG database, made it possible to produce a three-dimensional model of the lithologies in the study area.The model of the subsoil, simplified for applied reasons, was described in hy-drostratigraphic terms: three different lithotypes were subjected to piezometric levels monitor-ing. Finally, the research generated a numerical hydrological level in a stationary regime. In general, this study demonstrates how a numerical hydrogeological model calibrated by piezo-metric monitoring data can support the construction of a geological model, discarding or con-firming certain hypotheses and suggesting other means of reconstructing sedimentary bodies.

The ever-growing field of materials with applications in the biomedical field holds great promise regarding the design and fabrication of devices with specific characteristics especially scaffolds with personalized geometry and architecture. The continuous technological development pushes the limits of innovation in obtaining adequate scaffolds and establishing their characteristics and performance. To this end, computed tomography (CT) proved to be a reliable, non-destructive, high-performance machine, enabling visualization and structure analysis at sub-micronic resolutions. CT allows both qualitative and quantitative data of the 3D model, offering an overall image of its specific architectural features as well as reliable numerical data for rigorous analyses. The precise engineering of scaffolds consists in the fabrication of objects with well-defined morphometric parameters (e.g.: shape, porosity, wall thickness), and in their performance validation through thorough control over their behavior (in situ visualization, degradation, new tissue formation, wear, etc.). This review is focused on the use of CT in biomaterial science with the aim of qualitatively and quantitatively assess the scaffolds’ features and in monitoring their behavior following in vivo or in vitro experiments. Furthermore, the paper presents the benefits and limitations regarding the employment of this technique when engineering materials with applications in the biomedical field.

A position sensing glove, called SmartScan, that creates a 3D virtual model of a real object is presented. The data from the glove is processed by a volume minimization algorithm to validate the position sensor data. This allows only data from the object’s surface to be retained. The data validation algorithm allows the user to progressively improve an image by repeatedly moving their hand over the object. In addition, the user can choose their own balance between feature resolution and invalid data rejection. The SmartScan glove is tested on a foot model and is shown to be robust against motion artifacts, and has a mean accuracy of 2.9 mm (compared to a 3D model generated from optical imaging) without calibration.

A major problem in the post-inversion geophysical interpretation is the extraction of geological information from inverted physical property models, which do not necessarily represent all underlying geological features. No matter how accurate the inversions are, each inverted physical property model is sensitive to limited aspects of subsurface geology and is insensitive to other geological features that are otherwise detectable with complementary physical property models. Therefore, specific parts of the geological model can be reconstructed from different physical property models. To show how this reconstruction works, we simulated a complex geological system that comprises an original layered earth model that has passed several geological deformations and alteration overprints. Linear combination of complex geological features comprised three physical property distributions: Electrical resistivity, induced polarization chargeability, and magnetic susceptibility models. This study proposes a multivariate feature extraction approach to extract information about the underlying geological features comprising the bulk physical properties. We evaluated our method in numerical simulations and compared three feature extraction algorithms to see the tolerance of each method to the geological artifacts and noises. We show that the fast-independent component analysis (fast-ICA) algorithm by negentropy maximization is a robust method in the geological feature extraction that can handle the added unknown geological noises. The post-inversion physical properties are also used to reconstruct the underlying geological sources. We show that the sharpness of the inverted images is an important constraint on the feature extraction process. Our method successfully separates geological features in multiple 3D physical property models. This methodology is reproducible for any number of lithologies and physical property combinations and can recover the latent geological features, including the background geological patterns from overprints of chemical alteration.

Tuning fork gyroscopes (TFGs) are promising for potential high-precision applications. This work proposes and experimentally demonstrates a novel high-Q dual mass tuning fork microelectro-mechanical system (MEMS) gyroscope utilizing three-dimensional (3D) packaging techniques. Except for two symmetrically-decoupled proof masses (PM) with synchronization structures, a symmetrically-decoupled lever structure is designed to force the antiparallel, antiphase drive-mode motion and basically eliminate the low-frequency spurious modes. The thermoelastic damping (TED) and anchor loss are greatly reduced by the linearly-coupled, momentum- and torque-balanced antiphase sense mode. Besides, a novel 3D packaging technique is used to realize high Q-factors. A composite substrate encapsulation cap, fabricated by through-silicon-via (TSV) and glass-in-silicon (GIS) reflow processes, is anodically bonded to the sensing structures at wafer scales. A self-developed control circuit is adopted to realize loop control and characterize gyro-scope performances. It is shown that a high-reliability electrical connection together with a high-air-impermeability package can be fulfilled with this 3D packaging technique. Furthermore, the Q-factors of the drive and sense modes reach up to 51947 and 49249, respectively. This TFG realizes a wide measurement range of ±1800° /s and a high resolution of 0.1° /s with a scale-factor nonlinearity 720 ppm after automatic mode-matching. Besides, the long-term zero-rate output (ZRO) drift can be effectively suppressed by temperature compensation, inducing a small angle random walk (ARW) of 0.923°/√h and a low bias instability (BI) of 9.270°/h.

The current paper is focused on the rapidly developing field of nano-/micro three-dimensional production of inorganic materials. The fabrication method includes laserlithography of hybrid organic-inorganic materials with subsequent heat treatment lead-ing to a variety of crystalline phases in 3D structures. In this work, it was examineda series of organometallic polymer precursors with different silicon (Si) and zirconium (Zr) molar ratios, ranging from 9:1 to 5:5, prepared via sol-gel method. All mixtureswere examined for perspective used in 3D laser by manufacturing by fabricating nano-and micro-feature sized structures. Their deformation and surface morphology wereevaluated depending on chemical composition and crystallographic phase. The appear-ance of a crystalline phase was proven using single-crystal X-ray diffraction analysis,which revealed a lower crystallization temperature for microstructures compared tobulk materials. Fabricated 3D objects retain a complex geometry without any distortion after heat treatment up to 1400^oC. Under the proper conditions, a zircon phase (ZrSiO₄ - a highly stable material) can be observed. In addition, the highest newrecord of achieved resolution below 60 nm has been reached. The proposed prepara-tion protocol can be used to manufacture micro/nano-devices with high precision andresistance to high temperature and aggressive environment.

TThe paper presents the results of research on the influence of the settings of lubrication and cooling system parameters (solenoid valve opening time and lubricant feed pressure in terms of its quantity) in order to select the optimal lubricating conditions and thus reduce the wear of the dies used in the first forging operation of the valve forging made of high-nickel steel. Based on the observation of lubrication in the industrial process, it was found that a significant part of the lubricant fails to reach the die cavity, reaching the outside of it, which causes die wear due to seizure resulting from adhesion of the forging material to the tool surface, as well as high lubricant consumption and dirt in the press chamber. The authors proposed their own mobile lubricating and cooling system, which allows for a wide range of adjustments and provided with automatic cleaning procedures of the entire system, unlike the fixed lubrication system used so far in the industrial process. First, tests were carried out in laboratory conditions to determine the highest wettability and the lubricant remaining inside the tool cavity. These tests determined the lubrication system parameter settings which ensured that the greatest amount of lubricant remains in the cold die cavity without the forging process. Then, to verify the obtained results, tests were carried out in the industrial process of hot die forging of valve forgings, for short production runs of up to 500 forgings. The results were compared with the measurement of changes in the geometry of tools and forgings based on 3D scanning and surface topography analysis with the use of SEM. For best results (the variant of the setting of the dose and the time of exposure to lubricant), the forging process was carried out with the use of a new tool, up to the maximum service life.

In this study, a novel task of printing speed optimization for continuous fiber composites is investigated. Using continuous fibers is an innovative approach to reinforce products made by fused filament fabrication (FFF) additive manufacturing (AM) technology. In the printing process of composites with continuous fibers, the printing speed is critical because of its significant effect on the geometric shape of the samples, especially their corners. During optimization in this research, continuous glass fiber (CGF) and polylactic acid (PLA) filaments were utilized as reinforcing phase and matrix, respectively, and were simultaneously fed into the extrusion-based polymer 3D printer to form PLA/CGF composites. The optimization was carried out by calculating the temperature changes of the deposited rasters in the presence and absence of fibers as a first step and then determining the special relationship between the printing speeds and rasters temperature changes. Finally, the optimal and the maximum printing speed was computed based on a hypothesis, which is proved by the results of high-quality printed composites with different geometric shapes.

This paper investigated building data from multispectral and single-photon Lidar systems. The multispectral datasets from the individual channels and fused channels were explored. The multispectral and single-photon Lidar data were compared across multiple aspects: the data acquisition geometry, number of echoes, intensity, density, resolution, data defects, noise level, and the absolute and relative accuracy. In addition, we explored the performance of the multispectral and single-photon data for roof plane detection for eight complex/stylish buildings to investigate the suitability of these data for 3D building reconstruction. The building data from the single-photon and multispectral Lidar systems were evaluated with respect to the reference building vector data with an accuracy of better than 5 cm. The advantages and disadvantages of both technologies and their applications in the urban building environment are discussed.

Many classic visual monocular SLAM systems have been developed over the past decades, however, most of them will fail when dynamic scenarios dominate. DM-SLAM is proposed for handling dynamic objects in environments based on ORB-SLAM. The article mainly concentrates on two aspects. Firstly, DLRSAC is proposed to extract static features from the dynamic scene based on awareness of nature difference between motion and static, which is integrated into initialization of DM-SLAM. Secondly, we design candidate map points selection mechanism based on neighborhood mutual exclusion to balance the accuracy of tracking camera pose and system robustness in motion scenes. Finally, we conduct experiments in the public dataset and compare DM-SLAM with ORB-SLAM. The experiments verify the superiority of the DM-SLAM.

Synthetic polymer-based gradient foams have considered as promising category of functionally graded materials with unique properties. In this study, the carbon dioxide (CO₂) foaming technology has used for PET-PEN (Polyethylene Terephthalate - Polyethylene Naphthalate) copolymer towards porous functional materials with thermal insulation with reasonable mechanical strength. Through scanning electron microscope based morphological characterization, a potential to fabricate gradient foam structures with micro-pores has identified. It has shown that variation of post-foaming temperature can tune the pore size distribution although the very high post-foaming temperature tends to cause structural instability. Thermal measurement data set the limits of operation, confirmed by simultaneous differential scanning calorimeter and thermo-gravimetric analysis. Mechanical stress and thermal conductivity also has measured to find rationale of thermal insulation with reasonable mechanical strength and to elucidate the actual 3D grid foam of copolymer.

Three-dimensional (3D) printing has allowed for production of geometrically complex 3D objects with extreme flexibility, which is currently undergoing rapid expansions in terms of materials, functionalities, as well as areas of application. When attempting to print 3D microstructures in glass, femtosecond laser induced chemical etching (FLICE) – which is a subtractive 3D printing technique – has proved itself a powerful approach. Here, we demonstrate fabrication of macro-scale 3D glass objects of large heights up to ~3.8 cm with an identical lateral and longitudinal spatial resolution of ~20 μm. The remarkable accomplishment is achieved by revealing an unexplored regime in the interaction of ultrafast laser pulses with fused silica which results in aberration-free focusing of the laser pulses deeply inside fused silica.

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.

Development of novel anti-cancer peptides requires a rapid screening process which can be accelerated by using appropriate in vitro tumor models. Breast carcinoma tissue is a three dimensional (3D) microenvironment, which contains a hypoxic center surrounded by dense proliferative tissue. Biochemical clues provided by such 3D cell mass cannot be recapitulated in conventional 2D culture systems. In this experiment, we evaluate the efficacy of the sandalwood peptide, cyclosaplin on established in vitro 3D silk breast cancer model using invasive MDA-MB-231 cell line. The anti-proliferative effect of the peptide on 3D silk tumor model is monitored by alamar blue assay, with conventional 2D culture as control. The proliferation rate, glucose consumed, LDH, and MMP-9 activity of Human breast cancer cells are higher in 3D constructs compared to 2D. A higher concentration of drug is required to achieve 50% cell death in 3D culture than 2D cultures. The cyclosaplin treated MDA-MB-231 cells showed significant decrease in MMP-9 activity in 3D constructs. Microscopic analysis revealed the formation of cell clusters evenly distributed in the scaffolds. The drug treated cells were less in number, smaller and showed unusual morphology. Overall, these findings indicate the role of cyclosaplin as a promising anti-cancer therapeutics.

The objective of this study is to preliminarily evaluate a new CT-biopsy guidance device, an origami needle guide. The device is created by laser cutting the structure from a sheet of cardboard, 3D printing two radiocontrast agent grids on to the surface and folding the structure into a rectangular prism with a viewing window. An abdominal imaging phantom was used to evaluate the device through CT imaging and the targeting of lesions for needle insertion. The lesion targeting trials resulted in a mean targeting error of 1.88 mm with a standard deviation of 0.73 mm. The device attaches to the patient and is rigid enough to adequately support standard biopsy needles, reducing the effect of gravity and the risk of laceration by the needles, making it potentially advantageous for biopsy of superficial lesions and lesions approached from a horizontal orientation. The device supports insertion of multiple needles at once, making it particularly suitable for composite ablation using multiple needles. Another advantage of the device is that it can guide off-axial needle insertion. The low-cost and disposability of the device make it well-suited for the minimally invasive image guided therapy environment.

The capacity of 3D printing (3DP) technologies to initiate speedy polymerization of solvent free resins accounts for their utility in the manufacturing of medical devices. Nonetheless, independent biological evaluation of 3D printed materials is recommended due to the unique parameters of the manufacturing process, which can influence their physical, chemical, and biological properties. In this study, E-Shell 450 material indicated for 3DP of hearing aid shells and inner ear devices was examined for biological safety using zebrafish bioassays adapted to OECD fish embryo test. In addition, the proprietary material was characterized for composition using headspace gas-chromatography mass spectrometry (GC-MS). To initiate test, newly fertilized zebrafish eggs were cultured on non-treated and ethanol-treated materials in glass petri dishes with ultrapure water, incubated at 28.5°C and assessed for developmental endpoints of toxicity at 24h interval until 96h. Data confirmed non-treated material was extremely toxic in bioassays within 24h whereas ethanol-treated material showed a relative lower toxicity possibly due to ethanoic-aqueous interactions as observed by GC-MS. With the current influx of 3D printing materials, users are urged to exercise caution. Operators must also take cognizance of the potential toxicity of the chemicals used in 3DP and implement safety measures to limit their exposure.

The application of solvent method in the preparation of polypropylene powders was investigated, including the influence of various technological conditions, such as the raw material melt index of polypropylene, processing temperature, dosage of the solvent and the packing pressure. The experimental data show that using solvent method can effectively produce polypropylene powder, and the optimal technological conditions are: melt index of polypropylene being about 17.8 g/10min, processing temperature being about 166.0 ℃, dosage of the solvent being about 0.067g/ml and the packing pressure being about 0.7MPa.Under this technological conditions, polypropylene powder with excellent sphericity could be prepared, the apparent density of powder can reach 0.40 g/cm3 and the prepared powder displayed a narrow size distribution with the mean size of about 42.7 µm.

Thermal infrared imagery provides temperature information on target objects, and has been widely applied in non-destructive testing. However, thermal infrared imagery is not always able to display detailed textures of inspected objects, which hampers the understanding of geometric entities consisting of temperature information. Although some commercial software has been developed for 3D thermal model displays, the software requires the use of expensive specific thermal infrared sensors. This study proposes a cost-effective method for 3D thermal model reconstruction based on image-based modeling. Two smart phones and a low-cost thermal infrared camera are employed to acquire visible images and thermal images, respectively, that are fused for 3D thermal model reconstruction. The experiment results demonstrate that the proposed method is able to effectively reconstruct a 3D thermal model which extremely approximates its corresponding entity. The total computation time for the 3D thermal model reconstruction is intensive while generating dense points required for the creation of a geometric entity. Future work will improve the efficiency of the proposed method in order to expand its potential applications to in-time monitoring.

As part of our research for new leads against human African trypanosomiasis (HAT), we report on a 3D-QSAR study for antitrypanosomal activity and cytotoxicity of aminosteroid-type alkaloids recently isolated from the African medicinal plant Holarrhena africana A. DC. (Apocynaceae), some of which are strong trypanocides against Trypanosoma brucei rhodesiense (Tbr) with low toxicity against mammalian cells. Fully optimized 3D molecular models of seventeen congeneric Holarrhena alkaloids were subjected to a comparative molecular field analysis (CoMFA). CoMFA models were obtained for both, the anti-Tbr and cytotoxic activity data. Model performance was assessed in terms of statistical characteristics (R², Q² and P² for partial least squares (PLS) regression, internal cross-validation (leave-one-out) and external predictions (test set), respectively, as well as the corresponding SDEP and F-values). With R²=0.99, Q²=0.83 and P²=0.79 for anti-Tbr activity and R²=0.94, Q²=0.64, P²=0.59 for cytotoxicity against L6 rat skeletal myoblasts, both models were of good internal and external predictive power. The regression coefficients of the models representing the most prominent steric and electrostatic effects on anti-Tbr and for L6 cytotoxic activity were translated into contour maps and analyzed visually, allowing suggestions for possible modification of the aminosteroids to further increase the antitrypanosomal potency and selectivity. Very interestingly, the 3D-QSAR model established with the Holarrhena alkaloids also applied to the antitrypanosomal activity of two aminocycloartane-type compounds recently isolated by our group from Buxus sempervirens L. (Buxaceae), which indicates that these structurally similar natural products share a common SAR and, possibly, mechanism of action with the Holarrhena steroids. This 3D-QSAR study has thus resulted in plausible structural explanations of the antitrypanosomal activity and selectivity of aminosteroid- and aminocycloartane-type alkaloids as an interesting new class of trypanocides and may represent a starting point for lead optimization.

Triple-negative breast cancer (TNBC) is an aggressive and highly metastatic type of tumor. TNBC is often enriched in tumor-infiltrating neutrophils (TINs), which support cancer growth in part by counteracting tumor-infiltrating lymphocytes (TILs). Prior studies identified the enhancer of zeste homolog 2 (EZH2) as a pro-tumor methyltransferase in primary and metastatic TNBC. We hypothesized that EZH2 inhibition in TNBC cells per se would exert anti-tumor activity by altering the tumor immune microenvironment. To test this hypothesis, we used CRISPR to generate EZH2-knockout (KO) and overexpressing lines from parent (wild-type -WT) 4T1 cells, an established murine TNBC model. In vitro, EZH2 KO and OE cells showed little change in replicative capacity and invasiveness, but marked changes in surface marker profile and cytokine/chemokine secretion compared to WT cells. On in vivo injection, EZH2 KO cells showed significantly reduced primary tumor growth and a 10-fold decrease in lung metastasis compared to WT cells, while EZH2 OE cells were unchanged. Compared to WT tumors, TIN:TIL ratios were greatly reduced in EZH2 KO tumors but unchanged in EZH2 OE tumors. Our findings show a tumor-intrinsic role of EZH2 in the 4T1 TNBC model in regulating TIN/TIL poise, primary tumor progression and lung metastasis in vivo.

The Epithelial- Mesenchymal Transition (EMT) is a biological phenomenon associated with explicit phenotypic and molecular changes in cellular traits. Unlike the earlier-held popular belief of it being a binary process, EMT is now thought of as a landscape including diverse hybrid E/M phenotypes manifested by varying degrees of the transition. These hybrid cells can co-express both epithelial and mesenchymal markers and/or functional traits, and can possess the property of collective cell migration, enhanced tumor-initiating ability, and immune/targeted therapy-evasive features, all of which are often associated with worse patient outcomes. These characteristics of the hybrid E/M cells have led to a surge in studies that map their biophysical and biochemical hallmarks that can be helpful in exploiting their therapeutic vulnerabilities. This review discusses recent advances made in investigating hybrid E/M phenotype(s) from diverse biophysical and biochemical aspects by integrating live cell-imaging, cellular morphology quantification and mathematical modeling, and highlights a set of questions that remain unanswered about the dynamics of hybrid E/M states.

Connexin 43 (Cx43) forms gap junctions that mediate the direct intercellular diffusion of ions and small molecules between adjacent cells. Cx43 displays both pro- and anti-tumorigenic properties, but the mechanisms underlying these characteristics are not fully understood. Tunneling nanotubes (TNTs) are long and thin membrane projections that connect cells, facilitating the exchange of not only small molecules, but also larger proteins, organelles, bacteria, and viruses. Typically, TNTs exhibit increased formation under conditions of cellular stress and are more prominent in cancer cells, where they are generally thought to be pro-metastatic and to provide growth and survival advantages. Cx43 has been described in TNTs, where it is thought to regulate small molecule diffusion through gap junctions. Here, we developed a high-fidelity CRISPR/Cas9 system to knockout (KO) Cx43. We found that loss of Cx43 expression was associated with significantly reduced TNT length and number in breast cancer cell lines. Notably, secreted factors present in conditioned medium stimulated TNTs more potently when derived from Cx43-expressing cells than from KO cells. Moreover, TNT formation was significantly induced by inhibition of several key cancer signaling pathways that both regulate Cx43 and are regulated by Cx43, including RhoA kinase (ROCK), protein kinase A (PKA), focal adhesion kinase (FAK), and p38. Intriguingly, drug-induced stimulation of TNTs was more potent in Cx43 KO cells than in wild-type cells. In conclusion, this work describes a novel non-canonical role for Cx43 in regulating TNTs, identifies key cancer signaling pathways that regulate TNTs in this setting, and provides mechanistic insight into a pro-tumorigenic role of Cx43 in cancer.

We have previously demonstrated that receptor-interacting serine threonine kinase 1 (RIPK1) is expressed in hair follicle and regulates hair cycle. RIPK1 inhibitors also accelerated the telogen-to-anagen transition and elongated the anagen period in the mouse model. Here, we first investigated the involvement of RIPK1 in alopecia areata (AA). mRNA and protein expression of RIPK1 was increased in the skin of AA mouse model. Single cell RNA sequencing and immunohistochemistry showed that RIPK1 is highly increased in dendritic cells (DCs) and CD8+ T cells. RIPK1 inhibitors (i.e., Necrostatin-1s and GSK2982772) delayed the onset of AA in mouse model, and reduced the DCs and CD8+ T cells in AA skin. RIPK1 inhibitors also increased the hair length in the mouse hair organ culture mimicking AA. Collectively, these results suggest that RIPK1 is involved in AA onset via modulating immune cells, and RIPK1 inhibitors could prevent AA onset.

Cell mechanics is a biophysical indicator of cell state, such as cancer metastasis, leukocyte activation, and cell cycle progression. Atomic force microscopy (AFM) is a widely used technique to measure cell mechanics, where the Young modulus of a cell is usually derived from the Hertz contact model. However, the Hertz model assumes that the cell is an elastic, isotropic, and homogeneous material and that the indentation is small compared to the cell size. These assumptions neglect the effects of the cytoskeleton, cell size and shape, and cell environment on cell deformation. In this study, we investigated the influence of cell size on the estimated Young’s modulus using liposomes as cell models. Liposomes were prepared with different sizes and filled with phosphate buffered saline (PBS) or hyaluronic acid (HA) to mimic the cytoplasm. AFM was used to obtain the force indentation curves and fit them to the Hertz model. We found that the larger the liposome, the lower the estimated Young’s modulus for both PBS-filled and HA-filled liposomes. This suggests that the Young modulus obtained from the Hertz model is not only a property of the cell material, but also depends on the cell dimensions. Therefore, when comparing or interpreting cell mechanics using the Hertz model, it is essential to account for cell size.

The currently available anti-cancer therapies, such as gamma-radiation and chemotherapeutic agents, induce cell death and cellular senescence not only in cancer cells but also in the adjacent normal tissue. New anti-tumor approaches focus on limiting the side effects on normal cells. In this frame, the potential anti-tumor properties of Pulsed Electromagnetic Fields (PEMFs) through irradiation of breast cancer epithelial cells (MCF-7 and MDA-MB-231) and normal fibroblasts (FF95) were investigated. PEMFs had a frequency of 8 Hz, full-square wave type, magnetic flux density of 0.011 T and applied twice daily for 5 days. Data collected showcases that PEMF application decreases the proliferation rate and viability of breast cancer cells while having the opposite effect on normal fibroblasts. Moreover, PEMF irradiation induced cell death and cellular senescence only in breast cancer cells without any effect in the non-cancerous cells. These findings suggest PEMF irradiation as a novel, non-invasive anti-cancer strategy in combination with senolytic drugs, to eliminate both cancer and the remaining senescent cells while simultaneously avoiding the side effects of the current treatments.

Treating relapsed and refractory diffuse large B-cell lymphoma is still challenging for clinicians, but the available CAR-T and bispecific antibodies revolutionized therapy. Autologous stem cell transplantation was the most effective treatment modality previously. The authors report data from a single center over ten years. The retrospective study included 116 patients. There were 53 relapsed, 39 primary refractory cases, 19 had CNS involvement, and 5 received primary consolidation transplants. The median duration of follow-up was 46 months. The median event-free survival was 75 months, and the median overall survival was 105 months for all cases. Five-year overall survival was 59%, and event-free survival was 54%. Pretreatment prognostic factors at diagnosis had no effect on the outcome of transplantation. The authors found no difference between survival in relapsed or refractory cases, and the number of salvage lines or the germinal center / activated B-cell type also did not influence the results. Complete metabolic response before transplant confirmed by 18FDG PET/CT strongly affected survival. The pretransplant creatinine and CRP levels significantly influenced the long-term outcome. The number of stem cells infused did not affect survival, but engraftment within nine days did result in better survival. These data support the finding that the response to salvage therapy did select a better prognostic group who may still benefit from autologous transplantation.

Viruses can induce the fusion of infected and neighboring cells, leading to the formation of syncytia. Cell-cell fusion is mediated by viral fusion proteins on the plasma membrane of infected cells that interact with cellular receptors on neighboring cells. Viruses use this mechanism to spread rapidly to adjacent cells or escape host immunity. For some viruses, syncytium formation is a hallmark of infection and a known pathogenicity factor. For others, the role of syncytium formation in viral dissemination and pathogenicity remains poorly understood. Human cytomegalovirus (HCMV) is an important cause of morbidity and mortality in transplant patients and the leading cause of congenital infections. Clinical HCMV isolates have broad cell tropism but differ in their ability to induce cell-cell fusions, and little is known about the molecular determinants. We developed a system to analyze HCMV glycoprotein B (gB) variants in a defined genetic background. HCMV strains TB40/E and TR were used as vectors to compare the fusogenicity of six gB variants from congenitally infected fetuses with those from three laboratory strains. Five of them conferred the ability to induce fusion of MRC-5 human embryonic lung fibroblasts to one or both backbone strains, as determined by a split GFP-luciferase reporter system. The same gB variants were not sufficient to induce syncytia in infected ARPE-19 epithelial cells, suggesting that additional factors are involved. The system described here allows a systematic comparison of the fusogenicity of viral envelope glycoproteins and may help to clarify whether fusion-promoting variants are associated with increased pathogenicity.

G‐quadruplexes (G4) are structures formed at the ends of the telomere, these are rich in guanines and were stabilized by molecules that bind to specific sites. TMPyP4 and Thymoquinone (TQ) are small molecules that bind to the G4, they have drawn attention because of their role as telomerase inhibitors. The aim of this study was to evaluate the effects of telomerase inhibitors on cellular proliferation, senescence, and death. Two cell lines LC‐HK2 (NSCLC) and RPE‐1 were treated with TMPyP4 (5μM) and TQ (10μM). Both inhibitors were effective in decreasing telomerase activity. TMPyP4 increased the percentage of cells with membrane damage associated with cell death and decreased the frequency of cells in the S‐phase. TMPyP4 changed the cell adhesion ability and modified the pattern of focal adhesion. TQ acted in a dose‐dependent manner, increasing the frequency of senescent cells, and inducing cell cycle arrest in the G1. In conclusion, the effects of both drugs on LC-HK2 and RPE-1 cell lines were different although both are telomerase inhibitors, because TMPyP4 decreased proteins of cell adhesion and TQ induces a decrease in cell viability.

Here, our data provide the first evidence for the existence of a previously unknown receptive system formed by novel DNA- and RNA-based receptors in eukaryotes. This system, named the TR-system, is capable of recognizing and generating a response to different environmental factors and has been shown to orchestrate major vital functions of fungi, mammalian cells, and plants.Recently, we discovered the existence of a similar regulatory system in prokaryotes. These DNA- and RNA-based receptors are localized outside of the membrane forming a type of a network around cells that respond to a variety of chemical, biological, and physical factors and enabled the TR-system to regulate major aspects of eukaryotic cell life as follows: growth, including reproduction and development of multicellular structures; sensitivity to temperature, geomagnetic field, UV, light, and hormones; interaction with viruses; gene expression, recognition and utilization of nutrients. The TR-system was also implicated in cell memory formation and was determined to be responsible for its maintenance and the forgetting of preceding events. This system is the most distant receptive and regulatory system of the cell that regulates interactions with the outer environment and governs the functions of other receptor-mediated signaling pathways.

Background: Biological processes are based on complex networks of cells and molecules. Single cell multi-OMICs is a new tool aiming to provide new incites in the complex network of events controlling the functionality of the cell.; Methods: Since single cell technologies provide many sample measurements, they are the ideal environment for the application of deep learning and machine learning approaches. An autoencoder (AE) is composed of an encoder and a decoder sub-model. AE are very powerful in data compression and noise removal. However, the decoder model remains a black box from which is impossible to depict the contribution of the single input elements. We have recently developed a new class of autoencoders, called Sparsely Connected Autoencoders (SCA), which have the advantage of providing a controlled association among the input layer and the decoder module. This new architecture has the benefit that the decoder model is no anymore a black box and it can be used to depict new biologically interesting features from single cell data; Results: In this paper, we show that SCA hidden layer can grab new information usually hidden in single cell data, like as providing clustering on meta-features difficult, i.e. transcription factors expression, or impossible, miRNA expression, to depict in single cell RNAseq data. Furthermore, a SCA representation of cell clusters has the advantage of simulating a conventional bulk RNAseq, which is a data transformation allowing the identification of similarity among independent experiments; Conclusions: In our opinion, SCA represent the bioinformatics version of a “Swiss Army knife” for the extraction of hidden knowledgeable features from single cell OMICs data.