ABSTRACT
Sensory analysis methodologies are performed in sensory booths designed to minimise external stimuli, lacking ecological validity. Immersive environments are used to introduce contextual cues, but there is a lack of studies using mixed reality systems. The main goal of this study was to evaluate an augmented virtuality (AV) system where participants are inserted into a virtual environment and evaluate a real product, being able to interact with both dimensions. A panel of 102 consumers evaluated five samples of commercial peach nectars in three sessions, each in a different environment: public food court, living room (AV environments), and laboratory (traditional sensory booth). Consumers rated overall liking, followed by open comments, and also answered an Engagement (EQ) and a Presence Questionnaire (PQ). The type of environment only affected hedonic discrimination among samples, with the laboratory setting being the only one with sample discrimination. Nonetheless, each sample was not evaluated differently across the different environments. Concerning engagement, the environment only significantly influenced the EQ's 'Affective Value' factor, being higher when using an AV system. The level of presence in the virtual environment was significantly higher in the public food court, being significantly correlated with the EQ factor scores.
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The evolution in the biomedical engineering field boosts innovative technologies, with microfluidic systems standing out as transformative tools in disease diagnosis, treatment, and monitoring. Numerical simulation has emerged as a tool of increasing importance for better understanding and predicting fluid-flow behavior in microscale devices. This review explores fabrication techniques and common materials of microfluidic devices, focusing on soft lithography and additive manufacturing. Microfluidic systems applications, including nucleic acid amplification and protein synthesis, as well as point-of-care diagnostics, DNA analysis, cell cultures, and organ-on-a-chip models (e.g., lung-, brain-, liver-, and tumor-on-a-chip), are discussed. Recent studies have applied computational tools such as ANSYS Fluent 2024 software to numerically simulate the flow behavior. Outside of the study cases, this work reports fundamental aspects of microfluidic simulations, including fluid flow, mass transport, mixing, and diffusion, and highlights the emergent field of organ-on-a-chip simulations. Additionally, it takes into account the application of geometries to improve the mixing of samples, as well as surface wettability modification. In conclusion, the present review summarizes the most relevant contributions of microfluidic systems and their numerical modeling to biomedical engineering.
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Polydimethylsiloxane (PDMS) has attracted great attention in various fields due to its excellent properties, but its inherent hydrophobicity presents challenges in many applications that require controlled wettability. The purpose of this review is to provide a comprehensive overview of some key strategies for modifying the wettability of PDMS surfaces by providing the main traditional methods for this modification and the results of altering the contact angle and other characteristics associated with this property. Four main technologies are discussed, namely, oxygen plasma treatment, surfactant addition, UV-ozone treatment, and the incorporation of nanomaterials, as these traditional methods are commonly selected due to the greater availability of information, their lower complexity compared to the new techniques, and the lower cost associated with them. Oxygen plasma treatment is a widely used method for improving the hydrophilicity of PDMS surfaces by introducing polar functional groups through oxidation reactions. The addition of surfactants provides a versatile method for altering the wettability of PDMS, where the selection and concentration of the surfactant play an important role in achieving the desired surface properties. UV-ozone treatment is an effective method for increasing the surface energy of PDMS, inducing oxidation, and generating hydrophilic functional groups. Furthermore, the incorporation of nanomaterials into PDMS matrices represents a promising route for modifying wettability, providing adjustable surface properties through controlled dispersion and interfacial interactions. The synergistic effect of nanomaterials, such as nanoparticles and nanotubes, helps to improve wetting behaviour and surface energy. The present review discusses recent advances of each technique and highlights their underlying mechanisms, advantages, and limitations. Additionally, promising trends and future prospects for surface modification of PDMS are discussed, and the importance of tailoring wettability for applications ranging from microfluidics to biomedical devices is highlighted. Traditional methods are often chosen to modify the wettability of the PDMS surface because they have more information available in the literature, are less complex than new techniques, and are also less expensive.
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Over the last decade, researchers have developed a variety of new analytical and clinical diagnostic devices. These devices are predominantly based on microfluidic technologies, where biological samples can be processed and manipulated for the collection and detection of important biomolecules. Polydimethylsiloxane (PDMS) is the most commonly used material in the fabrication of these microfluidic devices. However, it has a hydrophobic nature (contact angle with water of 110°), leading to poor wetting behavior and issues related to the mixing of fluids, difficulties in obtaining uniform coatings, and reduced efficiency in processes such as plasma separation and molecule detection (protein adsorption). This work aimed to consider the fabrication aspects of PDMS microfluidic devices for biological applications, such as surface modification methods. Therefore, we studied and characterized two methods for obtaining hydrophilic PDMS surfaces: surface modification by bulk mixture and the surface immersion method. To modify the PDMS surface properties, three different surfactants were used in both methods (Pluronic® F127, polyethylene glycol (PEG), and polyethylene oxide (PEO)) at different percentages. Water contact angle (WCA) measurements were performed to evaluate the surface wettability. Additionally, capillary flow studies were performed with microchannel molds, which were produced using stereolithography combined with PDMS double casting and replica molding procedures. A PDMS microfluidic device for blood plasma separation was also fabricated by soft lithography with PDMS modified by PEO surfactant at 2.5% (v/v), which proved to be the best method for making the PDMS hydrophilic, as the WCA was lower than 50° for several days without compromising the PDMS's optical properties. Thus, this study indicates that PDMS surface modification shows great potential for enhancing blood plasma separation efficiency in microfluidic devices, as it facilitates fluid flow, reduces cell aggregations and the trapping of air bubbles, and achieves higher levels of sample purity.
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OBJECTIVE: The Ecological Momentary Assessment (EMA) concept was the basis for developing a smartphone application for the real-time report of awake bruxism (AB) activities. The aim of this study was to assess the effects of the EMA with a randomised controlled trial designed to compare the frequency of AB reported in two separate single observation points between a group of students monitored over time with an EMA smartphone application and a non-EMA monitored group. METHODS: Sixty-four (N = 64) dental students (22.3 YO [±2.6]; 52F; 12 M) answered the Oral Behaviour Checklist (OBC-1) and were randomised into an EMA-group (EG) and control group (CG). EG were monitored with a smartphone application for AB report during seven consecutive days and completed three EMA monitoring periods (EMA-1, 2 and 3) at one-month intervals. After the third period (EMA-3), both EG and CG answered again the OBC (OBC-2). Descriptive statistics were performed to characterise the sample. Pearson's chi-squared test, Fixer Exact test and post hoc Z test with Bonferroni correction were performed. The study hypothesis was that EMA could influence the AB reported by the participants because it implies an increase in the awareness, self-recognition, and self-control of bruxism behaviours. The level of significance was set at p < .05. RESULT: The percentage of participants who changed the report of the activity of clenching their teeth while awake between OBC-1 and OBC-2 was significantly higher among participants in EG, when compared to CG (p = .027). For all other behaviours reported the within- and between-group differences from OBC-1 to OBC-2 were not significant. Clenching and bracing were the most frequently reported, whilst grinding was the less frequent behaviour. CONCLUSION: A smartphone-based EMA approach demonstrated be useful to monitor AB over time, increasing individual's awareness, particularly to bracing and clenching behaviours.
Subject(s)
Bruxism , Ecological Momentary Assessment , Mobile Applications , Smartphone , Students, Dental , Humans , Male , Female , Students, Dental/psychology , Young Adult , Wakefulness/physiology , AdultABSTRACT
BACKGROUND AIMS: Parkinson's disease (PD) is the second most common neurodegenerative disorder. The etiology of the disease remains largely unknown, but evidence have suggested that the overexpression and aggregation of alpha-synuclein (α-syn) play key roles in the pathogenesis and progression of PD. Mesenchymal stromal cells (MSCs) have been earning attention in this field, mainly due to their paracrine capacity. The bioactive molecules secreted by MSCs, i.e. their secretome, have been associated with enhanced neuronal survival as well as a strong modulatory capacity of the microenvironments where the disease develops. The selection of the appropriate animal model is crucial in studies of efficacy assessment. Given the involvement of α-syn in the pathogenesis of PD, the evidence generated from the use of animal models that develop a pathologic phenotype due to the action of this protein is extremely valuable. Therefore, in this work, we established an animal model based on the viral vector-mediated overexpression of A53T α-syn and studied the impact of the secretome of bone marrow mesenchymal stromal cells MSC(M) as a therapeutic strategy. METHODS: Adult male rats were subjected to α-syn over expression in the nigrostriatal pathway to model dopaminergic neurodegeneration. The impact of locally administered secretome treatment from MSC(M) was studied. Motor impairments were assessed throughout the study coupled with whole-region (striatum and substantia nigra) confocal microscopy evaluation of histopathological changes associated with dopaminergic neurodegeneration and glial cell reactivity. RESULTS: Ten weeks after lesion induction, the animals received secretome injections in the substantia nigra pars compacta (SNpc) and striatum (STR). The secretome used was produced from bone marrow mesenchymal stromal cells MSC(M) expanded in a spinner flask (SP) system. Nine weeks later, animals that received the viral vector containing the gene for A53T α-syn and treated with vehicle (Neurobasal-A medium) presented dopaminergic cell loss in the SNpc and denervation in the STR. The treatment with secretome significantly reduced the levels of α-syn in the SNpc and protected the dopaminergic neurons (DAn) within the SNpc and STR. CONCLUSIONS: Our results are aligned with previous studies in both α-syn Caenorhabditis elegans models, as well as 6-OHDA rodent model, revealing that secretome exerted a neuroprotective effect. Moreover, these effects were associated with a modulation of microglial reactivity supporting an immunomodulatory role for the factors contained within the secretome. This further supports the development of new studies exploring the effects and the mechanism of action of secretome from MSC(M) against α-syn-induced neurotoxicity.
Subject(s)
Disease Models, Animal , Mesenchymal Stem Cells , Microglia , Neuroprotection , Parkinson Disease , alpha-Synuclein , Animals , Mesenchymal Stem Cells/metabolism , alpha-Synuclein/metabolism , alpha-Synuclein/genetics , Rats , Male , Microglia/metabolism , Parkinson Disease/therapy , Parkinson Disease/metabolism , Secretome/metabolism , Dopaminergic Neurons/metabolism , Mesenchymal Stem Cell Transplantation/methods , Cells, Cultured , HumansABSTRACT
Introduction: The inflammatory response after spinal cord injury (SCI) is an important contributor to secondary damage. Infiltrating macrophages can acquire a spectrum of activation states, however, the microenvironment at the SCI site favors macrophage polarization into a pro-inflammatory phenotype, which is one of the reasons why macrophage transplantation has failed. Methods: In this study, we investigated the therapeutic potential of the macrophage secretome for SCI recovery. We investigated the effect of the secretome in vitro using peripheral and CNS-derived neurons and human neural stem cells. Moreover, we perform a pre-clinical trial using a SCI compression mice model and analyzed the recovery of motor, sensory and autonomic functions. Instead of transplanting the cells, we injected the paracrine factors and extracellular vesicles that they secrete, avoiding the loss of the phenotype of the transplanted cells due to local environmental cues. Results: We demonstrated that different macrophage phenotypes have a distinct effect on neuronal growth and survival, namely, the alternative activation with IL-10 and TGF-ß1 (M(IL-10+TGF-ß1)) promotes significant axonal regeneration. We also observed that systemic injection of soluble factors and extracellular vesicles derived from M(IL-10+TGF-ß1) macrophages promotes significant functional recovery after compressive SCI and leads to higher survival of spinal cord neurons. Additionally, the M(IL-10+TGF-ß1) secretome supported the recovery of bladder function and decreased microglial activation, astrogliosis and fibrotic scar in the spinal cord. Proteomic analysis of the M(IL-10+TGF-ß1)-derived secretome identified clusters of proteins involved in axon extension, dendritic spine maintenance, cell polarity establishment, and regulation of astrocytic activation. Discussion: Overall, our results demonstrated that macrophages-derived soluble factors and extracellular vesicles might be a promising therapy for SCI with possible clinical applications.
Subject(s)
Interleukin-10 , Spinal Cord Injuries , Humans , Animals , Mice , Transforming Growth Factor beta1 , Proteomics , Secretome , Spinal Cord Injuries/therapyABSTRACT
This review attempts to provide a comprehensive assessment of recent methodologies, structures, and devices for pool boiling heat transfer enhancement. Several enhancement approaches relating to the underlying fluid route and the capability to eliminate incipient boiling hysteresis, augment the nucleate boiling heat transfer coefficient, and improve the critical heat flux are assessed. Hence, this study addresses the most relevant issues related to active and passive enhancement techniques and compound enhancement schemes. Passive heat transfer enhancement techniques encompass multiscale surface modification of the heating surface, such as modification with nanoparticles, tunnels, grooves, porous coatings, and enhanced nanostructured surfaces. Also, there are already studies on the employment of a wide range of passive enhancement techniques, like displaced enhancement, swirl flow aids, and bi-thermally conductive surfaces. Moreover, the combined usage of two or more enhancement techniques, commonly known as compound enhancement approaches, is also addressed in this survey. Additionally, the present work highlights the existing scarcity of sufficiently large available databases for a given enhancement methodology regarding the influencing factors derived from the implementation of innovative thermal management systems for temperature-sensitive electronic and power devices, for instance, material, morphology, relative positioning and orientation of the boiling surface, and nucleate boiling heat transfer enhancement pattern and scale. Such scarcity means the available findings are not totally accurate and suitable for the design and implementation of new thermal management systems. The analysis of more than 100 studies in this field shows that all such improvement methodologies aim to enhance the nucleate boiling heat transfer parameters of the critical heat flux and nucleate heat transfer coefficient in pool boiling scenarios. Finally, diverse challenges and prospects for further studies are also pointed out, aimed at developing important in-depth knowledge of the underlying enhancement mechanisms of such techniques.
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A computational fluid dynamics (CFD) model of blood flow through hyperbolic contraction with a discrete phase model (DPM) was experimentally validated. For this purpose, the positions and velocities of red blood cells (RBCs) flowing in a microchannel with hyperbolic contraction were experimentally assessed using image analysis techniques, and were subsequently compared with the numerical results. The numerically and experimentally obtained velocity fields were in good agreement, with errors smaller than 10%. Additionally, a nearly constant strain rate was observed in the contraction region, which can be attributed to the quasilinear increase in the velocity along the hyperbolic contraction. Therefore, the numerical technique used was validated due to the close similarity between the numerically and experimentally obtained results. The tested CFD model can be used to optimize the microchannel design by minimizing the need to fabricate prototypes and evaluate them experimentally.
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The examination of hyperelastic materials' behavior, such as polydimethylsiloxane (PDMS), is crucial for applications in areas as biomedicine and electronics. However, the limitations of hyperelastic models for specific stress scenarios, with stress concentration, are not well explored on the literature. To address this, firstly, three constitutive models were evaluated (Neo-Hookean, Mooney-Rivlin, and Ogden) using numerical simulations and Digital Image Correlation (DIC) analysis during a uniaxial tensile test. The samples were made of PDMS with stress concentration geometries (center holes, shoulder fillets, and edge notches). Results of ANOVA analysis showed that any of the three models can be chosen for numerical analysis of PDMS since no significant differences in suitability were found. Finally, the Ogen model was chosen to obtain the stress concentration factors for these geometries, a property which characterize how discontinuities change the maximum stress supported by an element. Our study provides new values for variables needed to analyze and design hyperelastic elements and produce a foundation for understanding PDMS stress-strain behavior.
Subject(s)
Dimethylpolysiloxanes , Stress, Mechanical , ElasticityABSTRACT
In this systematic review, the antimicrobial effect of ion-substituted calcium phosphate biomaterials was quantitatively assessed. The literature was systematically searched up to the 6th of December 2021. Study selection and data extraction was performed in duplo by two independent reviewers with a modified version of the OHAT tool for risk of bias assessment. Any differences were resolved by consensus or by a referee. A mixed effects model was used to investigate the relation between the degree of ionic substitution and bacterial reduction. Of 1016 identified studies, 108 were included in the analysis. The methodological quality of included studies ranged from 6 to 16 out of 18 (average 11.4). Selenite, copper, zinc, rubidium, gadolinium, silver and samarium had a clear antimicrobial effect, with a log reduction in bacteria count of 0.23, 1.8, 2.1, 3.6, 5.8, 7.4 and 10 per atomic% of substitution, respectively. There was considerable between-study variation, which could partially be explained by differences in material formulation, study quality and microbial strain. Future research should focus on clinically relevant scenarios in vitro and the translation to in vivo prevention of PJI.
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Introducción y objetivo: La pérdida masiva de peso provoca múltiples deformidades en el contorno corporal, incluida la región de los glúteos: pérdida de volumen y proyección, exceso de piel y ptosis tisular generalizada que dan lugar a antiestéticas deformidades. La reconstrucción quirúrgica de la región glútea en estos pacientes mediante lipoinjertos de grasa, colgajos adipocutáneos o colocación de implantes glúteos, arroja resultados subóptimos. El objetivo de este trabajo es presentar la versatilidad del colgajo de transposición adipomuscular de glúteo mayor en este tipo de deformidad. Material y método: Realizamos 20 remodelaciones glúteas con colgajo de transposición adipomuscular del glúteo mayor. Resultados: Los 20 pacientes fueron mujeres, con edades entre 33 y 61 años (media de 45 años). El colgajo de transposición adipomuscular de glúteo mayor se utilizó en combinación con lifting inferior circunferencial en 15 casos (75%), con lifting de glúteos escisional en 4 (20%) y en 1 caso secundario de deformidad provocada por colocación de implantes glúteos (5%). Conclusiones: En nuestra práctica clínica, el colgajo de transposición adipomuscular del glúteo mayor permite obtener buenos resultados estéticos en las deformidades glúteas posteriores a la pérdida masiva de peso en diferentes escenarios clínicos y en combinación con otros procedimientos, como el lifting circunferencial inferior o el escisional. También se puede realizar en procedimientos secundarios tras resultados insatisfactorios con la colocación de implantes glúteos. (AU)
Background and objective: Massive weight loss causes multiple body contour deformities, including the gluteal region. Volume and projection loss, excess skin associated with tissue ptosis lead to unsightly deformities of the gluteal contour. Surgical reconstruction of the gluteal region in these patients through fatgrafts, adipocutaneous flaps or gluteal implants have shown suboptimal results. The goal of this work is to present the versality of the transposition gluteus adipomuscular flap in these type of deformities. Methods: Twenty gluteal remodeling surgeries were performed using the transposition gluteus adipomuscular flap. Results: The 20 patients were women, aged between 33 and 61 years (mean 45 years). The transposition gluteus maximus adipomuscular flap was used in combination with lower bodylift in 15 cases (75%), with excisional gluteal lifting in 4 (20%) and in 1 secondary case of deformity caused by gluteal implant placement (5%). Conclusions: In our practice, the transposition gluteus adipomuscular flap allows good aesthetic results to be obtained in gluteal deformities after massive weight loss, namely in different clinical scenarios in combination with other procedures, such as the lower bodylift or the excisional gluteal lifting. It can also be performed in secondary procedures after unsatisfactory results using gluteal implants. (AU)
Subject(s)
Humans , Female , Adult , Middle Aged , Buttocks/surgery , Buttocks/abnormalities , Weight Loss , Surgical FlapsABSTRACT
The failure of axons to regenerate after a spinal cord injury (SCI) remains one of the greatest challenges in neuroscience. The initial mechanical trauma is followed by a secondary injury cascade, creating a hostile microenvironment, which not only is not permissive to regeneration but also leads to further damage. One of the most promising approaches for promoting axonal regeneration is to maintain the levels of cyclic adenosine monophosphate (cAMP), specifically by a phosphodiesterase-4 (PDE4) inhibitor expressed in neural tissues. Therefore, in our study, we evaluated the therapeutic effect of an FDA-approved PDE4 inhibitor, Roflumilast (Rof), in a thoracic contusion rat model. Results indicate that the treatment was effective in promoting functional recovery. Rof-treated animals showed improvements in both gross and fine motor function. Eight weeks post-injury, the animals significantly recovered by achieving occasional weight-supported plantar steps. Histological assessment revealed a significant decrease in cavity size, less reactive microglia, as well as higher axonal regeneration in treated animals. Molecular analysis revealed that IL-10 and IL-13 levels, as well as VEGF, were increased in the serum of Rof-treated animals. Overall, Roflumilast promotes functional recovery and supports neuroregeneration in a severe thoracic contusion injury model and may be important in SCI treatment.
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Kinship analysis from very low-coverage ancient sequences has been possible up to the second degree with large uncertainties. We propose a new, accurate, and fast method, correctKin, to estimate the kinship coefficient and the confidence interval using low-coverage ancient data. We perform simulations and also validate correctKin on experimental modern and ancient data with widely different genome coverages (0.12×-11.9×) using samples with known family relations and known/unknown population structure. Based on our results, correctKin allows for the reliable identification of relatedness up to the 4th degree from variable/low-coverage ancient or badly degraded forensic whole genome sequencing data.
Subject(s)
Forensic Medicine , Genome, Human , Humans , Whole Genome SequencingABSTRACT
Late diagnosis and systemic toxicity associated with conventional treatments make oncological therapy significantly difficult. In this context, nanomedicine emerges as a new approach in the prevention, diagnosis and treatment of cancer. In this work, pH-sensitive solid magnetoliposomes (SMLs) were developed for controlled release of the chemotherapeutic drug doxorubicin (DOX). Shape anisotropic magnetic nanoparticles of magnesium ferrite with partial substitution by calcium (Mg0.75Ca0.25Fe2O4) were synthesized, with and without calcination, and their structural, morphological and magnetic properties were investigated. Their superparamagnetic properties were evaluated and heating capabilities proven, either by exposure to an alternating magnetic field (AMF) (magnetic hyperthermia) or by irradiation with near-infrared (NIR) light (photothermia). The Mg0.75Ca0.25Fe2O4 calcined nanoparticles were selected to integrate the SMLs, surrounded by a lipid bilayer of DOPE:Ch:CHEMS (45:45:10). DOX was encapsulated in the nanosystems with an efficiency above 98%. DOX release assays showed a much more efficient release of the drug at pH = 5 compared to the release kinetics at physiological pH. By subjecting tumor cells to DOX-loaded SMLs, cell viability was significantly reduced, confirming that they can release the encapsulated drug. These results point to the development of efficient pH-sensitive nanocarriers, suitable for a synergistic action in cancer therapy with magnetic targeting, stimulus-controlled drug delivery and dual hyperthermia (magnetic and plasmonic) therapy.
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Microneedles (MNs) have been widely used in biomedical applications for drug delivery and biomarker detection purposes. Furthermore, MNs can also be used as a stand-alone tool to be combined with microfluidic devices. For that purpose, lab- or organ-on-a-chip are being developed. This systematic review aims to summarize the most recent progress in these emerging systems, to identify their advantages and limitations, and discuss promising potential applications of MNs in microfluidics. Therefore, three databases were used to search papers of interest, and their selection was made following the guidelines for systematic reviews proposed by PRISMA. In the selected studies, the MNs type, fabrication strategy, materials, and function/application were evaluated. The literature reviewed showed that although the use of MNs for lab-on-a-chip has been more explored than for organ-on-a-chip, some recent studies have explored this applicability with great potential for the monitoring of organ models. Overall, it is shown that the presence of MNs in advanced microfluidic devices can simplify drug delivery and microinjection, as well as fluid extraction for biomarker detection by using integrated biosensors, which is a promising tool to precisely monitor, in real-time, different kinds of biomarkers in lab- and organ-on-a-chip platforms.
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Adipose tissue-derived stem cells (ASCs) have been shown to assist regenerative processes after spinal cord injury (SCI) through their secretome, which promotes several regenerative mechanisms, such as inducing axonal growth, reducing inflammation, promoting cell survival, and vascular remodeling, thus ultimately leading to functional recovery. However, while systemic delivery (e.g., i.v. [intravenous]) may cause off-target effects in different organs, the local administration has low efficiency due to fast clearance by body fluids. Herein, a delivery system for human ASCs secretome based on a hydrogel formed of star-shaped poly(ethylene glycol) (starPEG) and the glycosaminoglycan heparin (Hep) that is suitable to continuously release pro-regenerative signaling mediators such as interleukin (IL)-4, IL-6, brain-derived neurotrophic factor, glial-cell neurotrophic factor, and beta-nerve growth factor over 10 days, is reported. The released secretome is shown to induce differentiation of human neural progenitor cells and neurite outgrowth in organotypic spinal cord slices. In a complete transection SCI rat model, the secretome-loaded hydrogel significantly improves motor function by reducing the percentage of ameboid microglia and systemically elevates levels of anti-inflammatory cytokines. Delivery of ASC-derived secretome from starPEG-Hep hydrogels may therefore offer unprecedented options for regenerative therapy of SCI.
Subject(s)
Neural Stem Cells , Spinal Cord Injuries , Rats , Humans , Animals , Glycosaminoglycans , Delayed-Action Preparations , Secretome , Spinal Cord Injuries/drug therapy , Heparin , Neural Stem Cells/metabolism , Spinal Cord , Adipose Tissue , Hydrogels , Polyethylene Glycols/metabolismABSTRACT
The ocean has a huge impact on our way of life; therefore, there is a need to monitor and protect its biodiversity [...].
Subject(s)
Biodiversity , Oceans and SeasABSTRACT
Symbiotic nitrogen fixation by Rhizobium bacteria in the cells of legume root nodules alleviates the need for nitrogen fertilizers. Nitrogen fixation requires the endosymbionts to differentiate into bacteroids which can be reversible or terminal. The latter is controlled by the plant, it is more beneficial and has evolved in multiple clades of the Leguminosae family. The plant effectors of terminal differentiation in inverted repeat-lacking clade legumes (IRLC) are nodule-specific cysteine-rich (NCR) peptides, which are absent in legumes such as soybean where there is no terminal differentiation of rhizobia. It was assumed that NCRs co-evolved with specific transcription factors, but our work demonstrates that expression of NCR genes does not require NCR-specific transcription factors. Introduction of the Medicago truncatula NCR169 gene under its own promoter into soybean roots resulted in its nodule-specific expression, leading to bacteroid changes associated with terminal differentiation. We identified two AT-Hook Motif Nuclear Localized (AHL) transcription factors from both M. truncatula and soybean nodules that bound to AT-rich sequences in the NCR169 promoter inducing its expression. Whereas mutation of NCR169 arrested bacteroid development at a late stage, the absence of MtAHL1 or MtAHL2 completely blocked bacteroid differentiation indicating that they also regulate other NCR genes required for the development of nitrogen-fixing nodules. Regulation of NCRs by orthologous transcription factors in non-IRLC legumes opens up the possibility of increasing the efficiency of nitrogen fixation in legumes lacking NCRs.
Subject(s)
Medicago truncatula , Rhizobium , Root Nodules, Plant/genetics , Medicago truncatula/genetics , Peptides/metabolism , Rhizobium/physiology , Glycine max/genetics , Gene Expression , Symbiosis/physiology , Gene Expression Regulation, PlantABSTRACT
Electroactive smart materials play an important role for tissue regenerative applications. Poly(vinylidene fluoride) (PVDF) is a specific subtype of piezoelectric electroactive material that generates electrical potential upon mechanical stimulation. This work focuses on the application of piezoelectric PVDF films for neural differentiation. Human neural precursor cells (hNPCs) are cultured on piezoelectric poled and non-poled ß-PVDF films with or without a pre-coating step of poly-d-lysine and laminin (PDL/L). Subsequently, hNPCs differentiation into the neuronal lineage is assessed (MAP2+ and DCX+ ) under static or dynamic (piezoelectric stimulation) culture conditions. The results demonstrate that poled and coated ß-PVDF films induce neuronal differentiation under static culture conditions which is further enhanced with mechanical stimulation. In silico calculations of the electrostatic potential of different domains of laminin, highlight the high polarity of those domains, which shows a clear preference to interact with the varying surface electric field of the piezoelectric material under mechanical stimulation. These interactions might explain the higher neuronal differentiation induced by poled ß-PVDF films pre-coated with PDL/L under dynamic conditions. Our results suggest that electromechanical stimuli, such as the ones induced by piezoelectric ß-PVDF films, are suitable to promote neuronal differentiation and hold great promise for the development of neuroregenerative therapies.