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Plant-specific transcription factors (TFs) are responsible for regulating the genes involved in the development of plant-specific organs and response systems for adaptation to terrestrial environments. This includes the development of efficient water transport systems, efficient reproductive organs, and the ability to withstand the effects of terrestrial factors, such as UV radiation, temperature fluctuations, and soil-related stress factors, and evolutionary advantages over land predators. In rice and Arabidopsis, INDETERMINATE DOMAIN (IDD) TFs are plant-specific TFs with crucial functions, such as development, reproduction, and stress response. However, in tomatoes, IDD TFs remain uncharacterized. Here, we examined the presence, distribution, structure, characteristics, and expression patterns of SlIDDs. Database searches, multiple alignments, and motif alignments suggested that 24 TFs were related to Arabidopsis IDDs. 18 IDDs had two characteristic C2H2 domains and two C2HC domains in their coding regions. Expression analyses suggest that some IDDs exhibit multi-stress responsive properties and can respond to specific stress conditions, while others can respond to multiple stress conditions in shoots and roots, either in a tissue-specific or universal manner. Moreover, co-expression database analyses suggested potential interaction partners within IDD family and other proteins. This study functionally characterized SlIDDs, which can be studied using molecular and bioinformatics methods for crop improvement.
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Arabidopsis , Solanum lycopersicum , Solanum lycopersicum/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , FilogeniaRESUMO
BACKGRUOUND: There is debate about ultrasonography screening for thyroid cancer and its cost-effectiveness. This study aimed to evaluate the cost-effectiveness of early screening (ES) versus symptomatic detection (SD) for differentiated thyroid cancer (DTC) in Korea. METHODS: A Markov decision analysis model was constructed to compare the cost-effectiveness of ES and SD. The model considered direct medical costs, health outcomes, and different diagnostic and treatment pathways. Input data were derived from literature and Korean population studies. Incremental cost-effectiveness ratio (ICER) was calculated. Willingness-to-pay (WTP) threshold was set at USD 100,000 or 20,000 per quality-adjusted life year (QALY) gained. Sensitivity analyses were conducted to address uncertainties of the model's variables. RESULTS: In a base case scenario with 50 years of follow-up, ES was found to be cost-effective compared to SD, with an ICER of $2,852 per QALY. With WTP set at $100,000, in the case with follow-up less than 10 years, the SD was cost-effective. Sensitivity analysis showed that variables such as lobectomy probability, age, mortality, and utility scores significantly influenced the ICER. Despite variations in costs and other factors, all ICER values remained below the WTP threshold. CONCLUSION: Findings of this study indicate that ES is a cost-effective strategy for DTC screening in the Korean medical system. Early detection and subsequent lobectomy contribute to the cost-effectiveness of ES, while SD at an advanced stage makes ES more cost-effective. Expected follow-up duration should be considered to determine an optimal strategy for DTC screening.
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Análise Custo-Benefício , Detecção Precoce de Câncer , Anos de Vida Ajustados por Qualidade de Vida , Neoplasias da Glândula Tireoide , Ultrassonografia , Humanos , Neoplasias da Glândula Tireoide/diagnóstico por imagem , Neoplasias da Glândula Tireoide/economia , Neoplasias da Glândula Tireoide/diagnóstico , República da Coreia/epidemiologia , Detecção Precoce de Câncer/economia , Detecção Precoce de Câncer/métodos , Ultrassonografia/economia , Ultrassonografia/métodos , Estudos Retrospectivos , Feminino , Masculino , Pessoa de Meia-Idade , Adulto , Cadeias de MarkovRESUMO
This study aimed to conduct importance-performance analyses (IPAs) based on Korean middle school students' health management awareness during the post-coronavirus disease 2019 (COVID-19) era. Data were collected from 867 Korean middle school students (13-15 years old) via online and offline surveys between May and June 2023. Frequency analysis, reliability analysis, IPA based on the entire student group, and IPA depending on sex were carried out with the collected data, which revealed the following. First, regardless of sex, the IPA results indicated that four factors of mental health were located in the third quadrant, with one factor of the same variable in the fourth quadrant. The three factors of disease management were located in the third quadrant. Regarding physical activity, two factors were located in the first quadrant, one in the second quadrant, and one in the third quadrant. Regarding sleep management, two factors were located in the second quadrant, one in the third quadrant, and one in the first quadrant. Regarding eating management, two factors were located in the third quadrant and one in the fourth quadrant. Regarding the social distancing variable, all four factors were located in the third quadrant. Regarding hygiene management, two factors were located in the first quadrant, one in the third quadrant, and one in the fourth quadrant. Furthermore, the IPA results indicated sex differences in regular sports and vigorous movement activities associated with physical activity. Additionally, a sex difference was observed in regular diet associated with eating management. This study proposed possible measures for encouraging middle school students to recognize the importance of health and increase their health-related performance during the COVID-19 endemic phase.
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Fluorescence-based bioimaging is an imperative approach with high clinical relevance in healthcare applications and biomedical research. The field of bioimaging plays an indispensable role in gaining insight into the internal architecture of cells/tissues and comprehending the physiological functions associated with biological systems. With the utility of piezoelectric nanomaterials, the bioelectric interface has been significantly investigated, leading to remarkable clinical relevance. Herein, we have developed barium titanate nanoparticle (BT) coated gold nanoclusters (AuNCs) in the presence and absence of an electromagnetic field (EMF). In this work, the effect of low (0.6 G) and high (2.0 G) EMFs on the structural arrangement of these piezoelectric nanocomposites (ABT) has been extensively studied with the help of X-ray diffraction (XRD), high diffraction resolution transmission electron microscopy (HR-TEM) and X-ray photoelectron spectroscopy (XPS). Furthermore, the two derivatives of ABT i.e. 0.6 ABT and 2.0 ABT have been evaluated for electrochemical behavior for their applicability as a candidate for exploring the bioelectric interface. Additionally, ABT, 0.6 ABT, and 2.0 ABT have been explored for cytocompatibility and bioimaging applications. The proposed piezoelectric nanocomposite, as a multifunctional platform, has enormous proficiency in the field of bioimaging and the capability to be utilized across the bioelectric interface.
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Nanocompostos , Nanopartículas , Bário , Ouro/química , Nanocompostos/químicaRESUMO
Although cannabidiol and tetrahydrocannabinol in Cannabis species exert their pharmacological effects via the endocannabinoid system, it is believed that other phytochemicals, particularly terpenes, can modulate therapeutic outcomes through the entourage effect. Therefore, to gain a better understanding of the pharmacological effects of Cannabis, obtaining information on phytochemical compositions, including mono-, di-, and sesqui-terpenes in Cannabis species is essential. Applying a sophisticated analytical method is indispensable. In this study, headspace-gas chromatography/mass spectrometry (HS-GC/MS) was employed to identify major terpenes in the leaves and inflorescences of hybrid Cannabis species. The incubation time and temperature conditions for HS-GC/MS were optimized. This method was successfully applied to the leaves (n = 9) and inflorescences (n = 7) of hybrid Cannabis species. A total of 26 terpenes in Cannabis species were detected, and six major components, such as α-pinene (9.8-2270 µg/g), ß-pinene (2.6-930 µg/g), myrcene (0.7-17,400 µg/g), limonene (1.3-300 µg/g), ß-caryophyllene (60-3300 µg/g), and α-humulene (40-870 µg/g), were quantified. Each sample showed different terpene compositions, but six major terpenes among all the terpenes detected were consistently found in both the leaves and inflorescences of hybrid Cannabis species. In this study, the six major terpenes' potential in hybrid Cannabis species was evaluated as biomarkers to distinguish hybrid Cannabis species samples. This study contributes to a better understanding of the entourage effect of Cannabis-based botanical drugs.
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Cannabis , Alucinógenos , Terpenos/análise , Cannabis/química , Inflorescência/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Limoneno/análise , Alucinógenos/análise , Agonistas de Receptores de Canabinoides , Compostos FitoquímicosRESUMO
Under severe environmental stress conditions, plants inhibit their growth and development and initiate various defense mechanisms to survive. The pseudo-response regulator (PRRs) genes have been known to be involved in fruit ripening and plant immunity in various plant species, but their role in responses to environmental stresses, especially high salinity and dehydration, remains unclear. Here, we focused on PRRs in tomato plants and identified two PRR2-like genes, SlSRP1 and SlSRP1H, from the leaves of salt-treated tomato plants. After exposure to dehydration and high-salt stresses, expression of SISRP1, but not SlSRP1H, was significantly induced in tomato leaves. Subcellular localization analysis showed that SlSRP1 was predominantly located in the nucleus, while SlSRP1H was equally distributed in the nucleus and cytoplasm. To further investigate the potential role of SlSRP1 in the osmotic stress response, we generated SISRP1-silenced tomato plants. Compared to control plants, SISRP1-silenced tomato plants exhibited enhanced tolerance to high salinity, as evidenced by a high accumulation of proline and reduced chlorosis, ion leakage, and lipid peroxidation. Moreover, SISRP1-silenced tomato plants showed dehydration-tolerant phenotypes with enhanced abscisic acid sensitivity and increased expression of stress-related genes, including SlRD29, SlAREB, and SlDREB2. Overall, our findings suggest that SlSRP1 negatively regulates the osmotic stress response.
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Desidratação , Solanum lycopersicum , Solanum lycopersicum/genética , Proteínas de Plantas/metabolismo , Cloreto de Sódio/farmacologia , Cloreto de Sódio/metabolismo , Ácido Abscísico/metabolismo , Estresse Fisiológico , Plantas Geneticamente Modificadas/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
Numerous staple crops exhibit polyploidy and are difficult to genetically modify. However, recent advances in genome sequencing and editing have enabled polyploid genome engineering. The hexaploid black nightshade species Solanum nigrum has immense potential as a beneficial food supplement. We assembled its genome at the scaffold level. After functional annotations, we identified homoeologous gene sets, with similar sequence and expression profiles, based on comparative analyses of orthologous genes with close diploid relatives Solanum americanum and S. lycopersicum. Using CRISPR-Cas9-mediated mutagenesis, we generated various mutation combinations in homoeologous genes. Multiple mutants showed quantitative phenotypic changes based on the genotype, resulting in a broad-spectrum effect on the quantitative traits of hexaploid S. nigrum. Furthermore, we successfully improved the fruit productivity of Boranong, an orphan cultivar of S. nigrum suggesting that engineering homoeologous genes could be useful for agricultural improvement of polyploid crops.
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Produtos Agrícolas , Poliploidia , Sequência de Bases , Mapeamento Cromossômico/métodos , Mutação , Fenótipo , Produtos Agrícolas/genética , Genoma de Planta/genética , Edição de GenesRESUMO
Various proteins introduced into living modified organism (LMO) crops function in plant defense mechanisms against target insect pests or herbicides. This study analyzed the antifungal effects of an introduced LMO protein, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium sp. strain CP4 (CP4-EPSPS). Pure recombinant CP4-EPSPS protein, expressed in Escherichia coli, inhibited the growth of human and plant fungal pathogens (Candida albicans, C. tropicalis, C. krusei, Colletotrichum gloeosporioides, Fusarium solani, F. graminearum, and Trichoderma virens), at minimum inhibitory concentrations (MICs) that ranged from 62.5 to 250 µg/mL. It inhibited fungal spore germination as well as cell proliferation on C. gloeosporioides. Rhodamine-labeled CP4-EPSPS accumulated on the fungal cell wall and within intracellular cytosol. In addition, the protein induced uptake of SYTOX Green into cells, but not into intracellular mitochondrial reactive oxygen species (ROS), indicating that its antifungal action was due to inducing the permeability of the fungal cell wall. Its antifungal action showed cell surface damage, as observed from fungal cell morphology. This study provided information on the effects of the LMO protein, EPSPS, on fungal growth.
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Antifúngicos , Fosfatos , Humanos , Antifúngicos/farmacologia , Plantas Geneticamente Modificadas/metabolismo , Fosfatos/farmacologia , 3-Fosfoshikimato 1-Carboxiviniltransferase/metabolismo , Fungos/metabolismo , Proteínas Recombinantes/farmacologia , Óxido Nítrico SintaseRESUMO
The characterization of an electron-positron beam generated from the interaction of a multi-GeV electron beam with a lead plate is performed using GEANT4 simulations. The dependence of the positron beam size on driver electron beam energy and lead converter thickness is investigated in detail. A pancake-like positron beam structure is generated with a monoenergetic multi-GeV driver electron beam, with the results indicating that a 5 GeV driver electron beam with 1 nC charge can generate a positron beam with a density of 1015-1016 cm-3 at one radiation length of lead. In addition, we find that electron-positron beams generated using above-GeV electron beams have neutralities greater than 0.3 at one radiation length of lead, whereas neutralities of 0.2 are observed when using a 200 MeV electron beam. The possibility of observing plasma instabilities in experiments is also examined by comparing the plasma skin depth with the electron-positron beam size. A quasi-neutral electron-positron plasma can be produced in the interaction between a 1 nC, 5 GeV electron beam and lead with a thickness of five radiation lengths. Our findings will aid in analyzing and interpreting laser-produced electron-positron plasma for laboratory astrophysics research.
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Forecasting the consequence of nanoparticles (NPs) and therapeutically significant molecules before materializing for human clinical trials is a mainstay for drug delivery and screening processes. One of the noteworthy obstacles that has prevented the clinical translation of NP-based drug delivery systems and novel drugs is the lack of effective preclinical platforms. As a revolutionary technology, the organ-on-a-chip (OOC), a coalition of microfluidics and tissue engineering, has surfaced as an alternative to orthodox screening platforms. OOC technology recapitulates the structural and physiological features of human organs along with intercommunications between tissues on a chip. The current review discusses the concept of microfluidics and confers cutting-edge fabrication processes for chip designing. We also outlined the advantages of microfluidics in analyzing NPs in terms of characterization, transport, and degradation in biological systems. The review further elaborates the scope and research on translational nanomedicines in human reproductive organs (testis, placenta, uterus, and menstrual cycle) by taking the advantages offered by microfluidics and shedding light on their potential future implications. Finally, we accentuate the existing challenges for clinical translation and scale-up dynamics for microfluidics chips and emphasize its future perspectives.
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Microfluídica , Nanopartículas , Humanos , Nanomedicina , Engenharia Tecidual , Sistemas MicrofisiológicosRESUMO
The development of the broad-bandwidth photon sources emitting in the soft X-ray range has attracted great attention for a long time due to the possible applications in high-resolution spectroscopy, nano-metrology, and material sciences. A high photon flux accompanied by a broad, smooth spectrum is favored for the applications such as near-edge X-ray absorption fine structure (NEXAFS), extended X-ray absorption fine structure (EXAFS), or XUV/X-ray coherence tomography (XCT). So far, either large-scale facilities or technologically challenging systems providing only limited photon flux in a single shot dominate the suitable sources. Here, we present a soft, broad-band (1.5 nm - 10.7 nm) soft X-ray source. The source is based on the interaction of very intense laser pulses with a target formed by a cluster mixture. A photon yield of 2.4 × 1014 photons/pulse into 4π (full space) was achieved with a medium containing Xe clusters of moderate-size mixed with a substantial amount of extremely large ones. It is shown that such a cluster mixture enhances the photon yield in the soft X-ray range by roughly one order of magnitude. The size of the resulting source is not beneficial (≤500 µm but this deficit is compensated by a specific spectral structure of its emission fulfilling the specific needs of the spectroscopic (broad spectrum and high signal dynamics) and metrological applications (broad and smoothed spectrum enabling a sub-nanometer resolution limit for XCT).
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Root hairs play crucial roles in the roots, including nutrient uptake, water assimilation, and anchorage with soil, along with supporting rhizospheric microorganisms. In rice, ammonia uptake is mediated by a specialized ammonium transporter (AMT). AMT1;1, AMT1;2, and AMT1;3 have been extensively studied in relation to nitrogen signaling. Cellulose synthase-like D1 (CSLD1) is essential for cell expansion and is highly specific to root hair cells. csld1 mutants showed successful initiation but failed to elongate. However, when nitrogen was depleted, csld1 root hairs resumed elongation. Further experiments revealed that in the presence of ammonium (NH4+), csld1 roots failed to elongate. csld1 elongated normally in the presence of nitrate (NO3−). Expression analysis showed an increase in root hair-specific AMT1;2 expression in csld1. CSLD1 was positively co-expressed with AMT1;2 changing nitrogen concentration in the growth media. CSLD1 showed increased expression in the presence of both ammonium and nitrate. Methylammonium (MeA) treatment of CSLD1 overexpression lines suggests that CSLD1 does not directly participate in nitrogen transport. Further studies on the root hair elongation mutant sndp1 showed that nitrogen assimilation is unlikely to depend on root hair length. Therefore, these results suggest that CSLD1 is closely involved in nitrogen-dependent root hair elongation and regulation of AMT1;2 expression in rice roots.
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In this study, we present an in situ microfluidic system to precisely control highly porous polycaprolactone microspheres as tissue templates for tissue engineering. The porosity of the microspheres was controlled by adjusting the flow rates of the polymer phase and the pore-generating material phase in the dispersed phase. The microfluidic flow-focusing technique was adopted to manufacture porous microspheres using a relatively highly viscous polymer solution, and the device was fabricated by conventional photolithography and PDMS casting. The fabricated in situ microfluidic system was used to precisely control the pore size of monodispersed polycaprolactone microspheres. The porous microspheres with controlled pore sizes were evaluated by culturing HDF cells on the surface of porous microspheres and injection into the subcutaneous tissue of rats. We found that the increased pore size of the microspheres improved the initial proliferation rate of HDF cells after seeding and relieved the inflammatory response after the implantation of porous microspheres in the subcutaneous tissue of rats.
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BACKGROUND: The purpose of this study was to establish a mechanical and histological basis for the development of biocompatible maxillofacial reconstruction implants by combining 3D-printed porous titanium structures and surface treatment. Improved osseointegration of 3D-printed titanium implants for reconstruction of maxillofacial segmental bone defect could be advantageous in not only quick osseointegration into the bone tissue but also in stabilizing the reconstruction. METHODS: Various macro-mesh titanium scaffolds were fabricated by 3D-printing. Human mesenchymal stem cells were used for cell attachment and proliferation assays. Osteogenic differentiation was confirmed by quantitative polymerase chain reaction analysis. The osseointegration rate was measured using micro computed tomography imaging and histological analysis. RESULTS: In three dimensional-printed scaffold, globular microparticle shape was observed regardless of structure or surface modification. Cell attachment and proliferation rates increased according to the internal mesh structure and surface modification. However, osteogenic differentiation in vitro and osseointegration in vivo revealed that non-mesh structure/non-surface modified scaffolds showed the most appropriate treatment effect. CONCLUSION: 3D-printed solid structure is the most suitable option for maxillofacial reconstruction. Various mesh structures reduced osteogenesis of the mesenchymal stem cells and osseointegration compared with that by the solid structure. Surface modification by microarc oxidation induced cell proliferation and increased the expression of some osteogenic genes partially; however, most of the markers revealed that the non-anodized solid scaffold was the most suitable for maxillofacial reconstruction.
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Implantes Dentários , Osseointegração , Humanos , Osteogênese , Impressão Tridimensional , Propriedades de Superfície , Titânio/química , Microtomografia por Raio-XRESUMO
Rice cultivation needs extensive amounts of water. Moreover, increased frequency of droughts and water scarcity has become a global concern for rice cultivation. Hence, optimization of water use is crucial for sustainable agriculture. Here, we characterized Loose Plant Architecture 1 (LPA1) in vasculature development, water transport, drought resistance, and grain yield. We performed genetic combination of lpa1 with semi-dwarf mutant to offer the optimum rice architecture for more efficient water use. LPA1 expressed in pre-vascular cells of leaf primordia regulates genes associated with carbohydrate metabolism and cell enlargement. Thus, it plays a role in metaxylem enlargement of the aerial organs. Narrow metaxylem of lpa1 exhibit leaves curling on sunny day and convey drought tolerance but reduce grain yield in mature plants. However, the genetic combination of lpa1 with semi-dwarf mutant (dep1-ko or d2) offer optimal water supply and drought resistance without impacting grain-filling rates. Our results show that water use, and transports can be genetically controlled by optimizing metaxylem vessel size and plant height, which may be utilized for enhancing drought tolerance and offers the potential solution to face the more frequent harsh climate condition in the future.
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During the coronavirus disease 2019 (COVID-19) pandemic, social distancing guidelines changed lifestyles, including increased sedentary time, physical inactivity, and disrupted sleep patterns among children. The purpose of the present study is to analyze the health awareness (mental health, disease, physical activity, sleep, eating habit, and hygiene health management) of elementary school students during the COVID-19 pandemic, and use the importance-performance analysis (IPA) technique to identify gender differences in health perceptions. We collected data on 1006 students, which was analyzed using frequency analysis, reliability testing, independent sample t-tests, and importance-performance analysis (IPA). A median importance value of 0.163 and a median performance value of 4.048 were selected as cross points to distribute the IPA matrix into four quadrants. The highest performance was given for wearing a mask and sanitary practice; the IPA matrix indicated that the sense of belonging, happiness, trust, and movement activity were located in quadrant I. Children's regular physical activity and level of physical activity were low, especially that of girls. Children's sleep management was poor. Their physical activity and sleep-related factors must be improved under the facilitation of the national government, public education institutions, and families.
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Papillary thyroid microcarcinoma (PTMC) has indolent features and low mortality. Recently, active surveillance (AS) instead of early surgery (ES) has been introduced as one treatment option but economical preference has not been established. The study objective was to systemically review the literature relating to cost-effectiveness of AS compared to ES for PTMC. Keywords were selected through PICO (Population, Intervention, Comparison, and Outcomes) tools. The search was conducted using PubMed, Cochrane, EMBASE, and Elsevier databases. Papers that had irrelevant titles were written in foreign languages, or had no original results were excluded. Out of the 62 papers extracted, five relevant to the subject matter of this study were identified. Three papers made their own decision models and proceeded with cost-effectiveness analysis (CEA), but the remaining two simply compared costs rather than cost-effectiveness. In terms of cost-effectiveness, three papers preferred AS, one preferred ES, and one preferred neither. The major differences in the CEA might arise from variations in each country's medical insurance system, the utility score systems, and decision models used. In subgroup analysis, two papers preferred AS to ES for patients at a younger age at diagnosis in terms of cost-effectiveness as well as tumor biological characteristics. Although AS has been generally more cost-effective than ES in previous publications, younger age at diagnosis could be one factor contributing to preference for ES. The CEA of prospective cohorts based on the decision model and utility score for thyroid cancer should be undertaken to confirm the cost-effectiveness of AS.
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The construction of an in vitro 3D cellular model to mimic the human liver is highly desired for drug discovery and clinical applications, such as patient-specific treatment and cell-based therapy in regenerative medicine. However, current bioprinting strategies are limited in their ability to generate multiple cell-laden microtissues with biomimetic structures. This study presents a method for producing hepatic-lobule-like microtissue spheroids using a bioprinting system incorporating a precursor cartridge and microfluidic emulsification system. The multiple cell-laden microtissue spheroids can be successfully generated at a speed of approximately 45 spheroids min-1 and with a uniform diameter. Hepatic and endothelial cells are patterned in a microtissue spheroid with the biomimetic structure of a liver lobule. The spheroids allow long-term culture with high cell viability, and the structural integrity is maintained longer than that of non-structured spheroids. Furthermore, structured spheroids show high MRP2, albumin, and CD31 expression levels. In addition, the in vivo study reveals that structured microtissue spheroids are stably engrafted. These results demonstrate that the method provides a valuable 3D structured microtissue spheroid model with lobule-like constructs and liver functions.
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Materiais Biomiméticos/química , Albuminas/genética , Albuminas/metabolismo , Animais , Materiais Biomiméticos/metabolismo , Bioimpressão , Sobrevivência Celular , Células Cultivadas , Células Endoteliais/metabolismo , Humanos , Dispositivos Lab-On-A-Chip , Fígado , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteína 2 Associada à Farmacorresistência Múltipla/genética , Proteína 2 Associada à Farmacorresistência Múltipla/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Esferoides Celulares/metabolismo , Engenharia TecidualRESUMO
With remarkable developments in technologies, the possibility of replacing injured tissue or organs with artificial ones via three-dimensional bioprinting is being improved. The basic prerequisite for successful application of bioprinting is high cell survival following printing. In this study, numerical calculations and experiments were performed to understand cell damage process incurred by forced extrusion bioprinters. Compressible and shear stresses were presumed to play a pivotal role within the syringe and needle, respectively, based on numerical calculation. To verify the numerical results, two experiments-pressurization in a clogged syringe and extrusion through syringe-needle-were conducted, and the damaged cell ratio (DCR) were measured by live/dead assays. Shear stress of needle flow had a great influence on DCR of discharged bioink, whereas effect of compressible stress in clogged syringe was relatively small. Cell damage in the needle flow is affected by moving distance under load as well as magnitude of shear stress. Applying this concept the differential equation of DCR growing was established, similar to the historied logistic equation for population dynamics, and the mathematical formula to predict DCR was explicitly represented splendidly as a function of only one independent variable, pressure work. The proposed formula was able to effectively predict DCR measurements for 43 bioprinting conditions, and the exactness confirmed the hypothesis for the theory. The presence of safe core zone, which may be related to the critical shear stress and stressed duration on cells, was theoretically conjectured from the DCR measurements, and further studies are necessary for an extensive and profound understanding. Fast printing is required for efficiency of a bio-structure fabrication; however, the higher shear stress accompanying increased operating pressure to speed up bioink discharge rate causes more cell damage. Employing the accurate formula presented, the optimal bioprinting conditions can be designed with ensuring targeted cell viability.
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Bioimpressão , Sobrevivência Celular , Impressão Tridimensional , Reologia , Estresse Mecânico , Engenharia Tecidual , Alicerces TeciduaisRESUMO
Ralstonia solanacearum causes bacterial wilt disease in solanaceous crops. Identification of avirulence type III-secreted effectors recognized by specific disease resistance proteins in host plant species is an important step toward developing durable resistance in crops. In the present study, we show that R. solanacearum effector RipJ functions as an avirulence determinant in Solanum pimpinellifolium LA2093. In all, 10 candidate avirulence effectors were shortlisted based on the effector repertoire comparison between avirulent Pe_9 and virulent Pe_1 strains. Infection assays with transgenic strain Pe_1 individually carrying a candidate avirulence effector from Pe_9 revealed that only RipJ elicits strong bacterial wilt resistance in S. pimpinellifolium LA2093. Furthermore, we identified that several RipJ natural variants do not induce bacterial wilt resistance in S. pimpinellifolium LA2093. RipJ belongs to the YopJ family of acetyltransferases. Our sequence analysis indicated the presence of partially conserved putative catalytic residues. Interestingly, the conserved amino acid residues in the acetyltransferase catalytic triad are not required for effector-triggered immunity. In addition, we show that RipJ does not autoacetylate its lysine residues. Our study reports the identification of the first R. solanacearum avirulence protein that triggers bacterial wilt resistance in tomato. We expect that our discovery of RipJ as an avirulence protein will accelerate the development of bacterial wilt-resistant tomato varieties in the future.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.