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The Living Wall (LW) garden system has been employed as a post-treatment system to improve the effluent quality of septic tanks. This improvement primarily involves reducing nutrient levels, as well as facilitating the removal of organic matter and solids in accordance with effluent discharge guidelines. The objective of this study was to investigate the treatment performance of the LW system connected to a septic tank, along with an examination of the microbial communities within the LW units. A laboratory-scale LW system, comprising LW1, LW2, and LW3 units, was employed. The system was fed with effluent obtained from septic tanks and varied by theoretical hydraulic retention time (HRT) of 6, 12, and 24 h. The TCOD, SCOD, TSS, TVS, TKN, and TP removal efficiencies of the LWs were achieved at 62 ± 24, 42 ± 19, 72 ± 21, 66 ± 15, 80 ± 15, and 58 ± 21%, respectively. To classify microbial communities in the soil and gravels collected from each LW unit, the Illumina MiSeq System Sequencer was employed. Nitrospirota was consistently found in all LW units, aiding in the conversion of nitrogen. Fusobacteriota were detected in specific layers of the LW units, indicating varying oxygen levels in the LW system.
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The effectiveness of phosphorus (P) removal by sand filters is limited during septic tank effluent (STE) treatment. The elevated effluent P concentrations pose threats to drinking water quality and contribute to eutrophication. The concern of P leaching from sand filters is further exacerbated by the increased frequency of flooding and natural precipitation due to climate change. This study aimed to understand P attenuation and leaching dynamics, as well as the removal mechanisms in sand filters treating STE, offering insights into the design and implementation of P removal/recovery modules to onsite wastewater treatment systems. P attenuation and leaching during STE treatment and rainfall were studied in bench-scale columns (new vs. aged sand). At standard STE loading (1.2 gallon d-1 ft-2), 24-32% removal of total phosphorus (TP) was achieved, while increased P removal efficiency (35-53%) was observed at low loading (0.6 gallon d-1 ft-2) with influent containing 10.3-20.0 mg P L-1. Complete breakthrough was observed in both aged (12-70 days) and new columns (27-73 days) at test hydraulic loadings. The maximum TP attenuation level was 20.6-45.3 mg P kg-1 and 25.3-33.0 mg P kg-1, in aged and new sand columns, respectively. When simulated rain was applied (15-60 mm h-1), 80-97% of the attenuated P leached out and the leaching dynamics were impacted by rainfall duration rather than the intensity. The highest concentrations of TP (15.6-15.9 mg L-1) were leached out from both columns within the first 2-6 hours. Orthophosphate was the dominant P species in treated effluent (83-84%) and leachate (69-88%), demonstrating its significance as the major P form in the discharge. In addition, aged sand (>5 years) accumulated higher levels of Mg, Al, Ca, and Fe, thus enhancing the P attenuation level during STE treatment. Collectively, this study underscored the importance of frequent field monitoring for reliable long-term P removal estimates.
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In this paper, the problem of highly performance motion control of tank bidirectional stabilizer with dead zone nonlinearity and uncertain nonlinearity is addressed. First, the electromechanical coupling dynamics model of bidirectional stabilizer is developed finely. Second, the dead zone nonlinearity in bidirectional stabilizer is characterized as the combination of an uncertain time-varying gain and a bounded disturbance term. Meanwhile, an adaptive robust controller with dead zone compensation is proposed by organically combining adaptive technique and extended state observer (ESO) through backstepping method. The adaptive technique is employed to reduce the impact of unknown system parameter and dead zone parameter. Furthermore, the ESO is constructed to compensate the lumped uncertainties including unmodeled dynamics and dead zone residual, and integrated together via a feedforward cancellation technique. Moreover, the adaptive robust control law is derived to ensure final global stability. In stability analysis, the asymptotic tracking performance of the proposed controller can be guaranteed as the uncertainty nonlinearities in tank bidirectional stabilizer are constant. It is also guaranteed to achieve bounded tracking performance when time-varying uncertainties exist. Extensive co-simulation and experimental results verify the superiority of the proposed strategy.
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Polyacrylonitrile (PAN) nanofibers have specific characteristics such as thermal insulation, weatherproofing, and sunlight resistance and therefore are appropriate to be applied as insulation materials for various industries, especially in greenhouse construction. The heat source in greenhouse buildings that operate independently in the heating network comes from heat storage tanks. In the present study, employing thermal field numerical simulations, we investigate the heat flux of a cylindrical heat storage tank with silica aerogel-modified PAN nanofibers as thermal insulation materials. The geometric scale of the tank body, thermal insulation material thickness, and outdoor temperature are optimized to improve thermal insulation. The significant discrepancy in heat flux at different parts of the heat storage tank leads to the extreme heat flux arising at the water-gas interface on the inner and outer walls. It is indicated that the heat flux distribution can be effectively ameliorated by modifying the scale of the tank body to retain the overall water temperature. In particular, effective insulation can merely be acquired when the thermal conductivity of the insulation material is below 3.3 W·m-1·K-1. Eventually, the heat storage tank is optimized to store 1400 L water at 100 °C with a radius of 0.6 m and a thermal insulation thickness of 50 mm at an outdoor temperature of -10 °C, which can maintain excellent thermal insulation for 8 and 24 h at 87.7 and 69.9 °C, respectively.
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The Baculovirus Expression Vector System (BEVS) has revolutionized the field of recombinant protein expression by enabling efficient and high yield production. The platform offers many advantages including manufacturing speed, flexible design, and scalability. In this chapter, we describe the methods including strategies and considerations to successfully optimize and scale-up using BEVS as a tool for production (Fig. 1). As an illustrative case study, we present an example focused on the production of a viral glycoprotein.
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Baculoviridae , Vetores Genéticos , Proteínas Recombinantes , Baculoviridae/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/biossíntese , Vetores Genéticos/genética , Animais , Humanos , Células Sf9RESUMO
The Nísia Floresta National Forest, located in Rio Grande do Norte (RN), is an important remnant of the Atlantic Forest biome in Brazil. Bromeliad tanks in this forest offer suitable breeding sites for mosquito species that may act as viral vectors, thus posing an epidemiological concern. However, studies investigating the presence of immature Culicidae in natural breeding sites in RN have thus far been restricted to Caatinga vegetation. This study investigated mosquitoes and their natural breeding sites in bromeliads growing in the Nísia Floresta National Forest. From March 2013 to February 2014, monthly samples were collected from the tanks of five randomly selected bromeliads and larvitraps placed in each of the three forest management areas. Hohenbergia catingae Ule (Hohenbergia bromeliad) is an important shelter for immature mosquitoes. Culex (Microculex) was the predominant species, representing 86% of the immature mosquitoes collected. A rare occurrence of Aedes (Stegomyia) aegypti (Linnaeus, 1762) (generally associated with urban areas under high anthropogenic influence) was observed, highlighting the importance of investigating the presence of mosquitoes in different natural habitats. An analysis of species diversity revealed that species such as Culex imitator Theobald, 1903 and Culex davisi Kumm, 1933, have a strong association with bromeliads. In tire traps (larvitraps) Aedes (Stegomyia) albopictus Skuse, 1894 was predominant. Environmental changes, such as deforestation, removal of bromeliads, and climate change in the area, can influence the migration of species and adaptation to new habitats in a peridomiciliary environment around the forest, consequently the possibility of transmission of virus and other pathogens.
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BACKGROUND: In epiphytic bromeliads, the roots used to be considered poorly functional organs in the processes of absorption and metabolization of water and nutrients, while the leaves always acted as protagonists in both functions. More recent discoveries have been changing this old view of the root system. SCOPE: In this review, we will address the old thoughts of the scientific community regarding the function performed by the roots of epiphytic bromeliads (mere holdfast structures with low physiological activity) and the importance of a reduced or lack of root system for the emergence of epiphytism. We will present indirect and direct evidence that contradicts this older hypothesis. Furthermore, the importance of the root absorptive function mainly for juvenile tankless epiphytic bromeliads and the characteristics of the root absorption process of adult epiphytic tank bromeliads will be thoroughly discussed in physiological aspects. Finally, some factors (species, substrate, environmental conditions) that influence the absorptive capability of the roots of epiphytic tank bromeliads will also be considered in this review, highlighting the importance that the absorptive role of the roots have for the plasticity of bromeliads that live on trees, which is an environment characterized by the intermittent availability of water and nutrients. CONCLUSIONS: The roots of tank-forming epiphytic bromeliads play important roles in the absorption and metabolization of nutrients and water. The importance of roots stands out mainly for juvenile tankless bromeliads since the root is the main absorptive organ. In larger plants with tank, although the leaves become the protagonists in the resource acquisition process, the roots complement the absorptive function of the leaf trichomes, resulting in a better growth of the bromeliad. The physiological and biochemical properties of the processes of absorption and distribution of resources in the tissues seem to differ between absorption by trichomes and roots.
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Chronic treatment with clomipramine, a tricyclic antidepressant drug, reduces symptoms of obsessive-compulsive disorder (OCD) and can influence the activity of the hypothalamic-pituitary-adrenal axis. However, little is known regarding the effects of acute clomipramine on the immediate expression of stress responses. Serotonergic drugs can elicit surfacing, a behavioral profile potentially related to toxicity in fish, although surfacing has not yet been observed after clomipramine exposure. The present study investigated the impact of acute exposure to clomipramine on basal and stress-induced behaviors in the novel tank test and cortisol levels in mixed-sex, wild-type, adult zebrafish (Danio rerio). The findings show clomipramine-exposed groups (regardless of stress exposure) spent much more time in the top of the novel tank and had significantly less overall motor activity in the behavioral task compared to the fish not exposed to the drug. Then, the dose-dependent effects of acute clomipramine on activity in the surface of the novel tank (top third of the top half) were investigated further. Clomipramine dose-dependently increased surface-dwelling and elicited a dose-dependent hypoactivity in overall motor behavior. There were no statistically significant differences in whole-body cortisol levels in either experiment. Like other serotonin-acting drugs, clomipramine strongly elicited surface-dwelling and depressed motor behavior in adult zebrafish. Additional testing is needed to elucidate whether surfacing represents a toxic state and how serotonin regulates surfacing.
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Comportamento Animal , Clomipramina , Relação Dose-Resposta a Droga , Hidrocortisona , Peixe-Zebra , Animais , Clomipramina/farmacologia , Clomipramina/administração & dosagem , Comportamento Animal/efeitos dos fármacos , Hidrocortisona/metabolismo , Masculino , Feminino , Antidepressivos Tricíclicos/farmacologia , Antidepressivos Tricíclicos/administração & dosagem , Inibidores Seletivos de Recaptação de Serotonina/farmacologia , Inibidores Seletivos de Recaptação de Serotonina/administração & dosagemRESUMO
This study investigated the effects of anxiogenic and anxiolytic drugs on zebrafish (Danio rerio) behaviour using a modified novel tank dive test with higher walls and a narrower depth. Zebrafish were administered chondroitin sulfate, beta-carboline, delta-9-tetrahydrocannabinol (THC), ethanol, and beta-caryophyllene, and their behaviours were evaluated for geotaxis, swimming velocity, and immobility. Both anxiogenic and anxiolytic compounds generally increased bottom-dwelling behaviour, suggesting that the tank's modified dimensions significantly influence zebrafish responses. EC50 values for ethanol showed a lower threshold for velocity reduction compared to zone preference. Chondroitin sulfate uniquely caused a sex-specific increase in male swimming velocity, whereas no other sex-differences were observed with any compound. Interestingly, the presence of drug-treated fish did not alter the behaviour of observer fish, suggesting limited social buffering effects. The findings underscore the complexity of zebrafish behavioural phenotypes and highlight the need for considering tank dimensions and multiple behavioural parameters to accurately assess the effects of anxiety-modulating drugs. This study demonstrates the utility of the modified novel tank dive test in providing nuanced insights into the behavioural effects of different pharmacological agents in zebrafish.
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In this study, we focused on the 3D dispersion of colloids. To our knowledge, we were the first to do so. Thereto, we injected silica encapsulated DNA tagged superparamagnetic particles (SiDNAmag) in a homogeneous coarse grain sand tank. At four downstream locations, SiDNAmag concentrations were determined as a function of time. Longitudinal and transverse dispersivity values and associated uncertainties of SiDNAmag were determined using Monte Carlo modelling approach. The parameter associated uncertainties of hydraulic conductivity as well as of the effective porosity estimated from SiDNAmag breakthrough curves were statistically similar to those estimated from salt tracer breakthrough curves. Further, the SiDNAmag dispersivity uncertainty ranges were then statistically compared with the salt tracer (NaCl, and fluorescein) dispersivities. Our results indicated that time to rise, time of peak concentration and shape of the breakthrough curves of SiDNAmag were similar to those of the salt tracer breakthrough curves. Despite the size difference between the salt tracer molecules and SiDNAmag, size exclusion did not occur, probably due to the large pore throat diameter to SiDNAmag diameter ratio. The median longitudinal dispersivity (αL) of salt tracer and SiDNAmag were 4.9 and 5.8 × 10-4 m, respectively. The median ratio of horizontal and vertical transverse dispersivities to αL, (αTH /αL and αTV /αL, respectively), for salt tracer and SiDNAmag ranged between 0.52 and 0.56. Through the statistical tests, we concluded that the longitudinal and traverse dispersivities of SiDNAmag were not statistically significantly different from salt tracer in 3 dimensions and could be used to characterize the dispersive properties of the medium we used. Our work contributes to a better understanding of 3D dispersion of SiDNAmag in saturated porous media.
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There is an increasing interest in the microbiota of the dairy value chain, from field to fork. Studies to understand the effects of environmental, feed and management factors on the raw milk microbiota have been performed to elucidate the origin of the bacteria and find ways to control the presence or absence of specific bacteria. In this study, we explored the microbiota in feedstuff, bedding material and milk on a Swedish dairy farm to investigate the effects of feeding different silages on the bacterial compositions throughout the dairy value chain. Three ensiling treatments were evaluated: without additive, with acid treatment, and with inoculation of starter culture. The silage treatments were fed as partial mixed rations to 67 dairy cows for 3 weeks each, with one treatment fed twice to evaluate if a potential change in milk microbiota could be repeated. The highest average total bacteria counts were found in the used bedding material (9.6 log10 cfu/g), while milk showed the lowest (3.5 log10 cfu/g). Principal coordinate analysis of the weighted UniFrac distance matrix showed clear separation between 3 clusters of materials: 1) herbage, 2) silage and partial mixed ration, and 3) used bedding material and milk. Surprisingly, the expected effect of the ensiling treatments on silage microbiota was not clear. Transfer of major bacteria from the silages and resulting partial mixed rations to the used bedding material was observed, but rarely to milk. The milk microbiota showed most resemblance to that of the used bedding material. Lactobacillus was a major genus in both feed and milk, but investigations at amplicon sequence variant level showed that in most cases the sequences differed between materials. However, low total bacteria count in the milk in combination with a high diversity suggests that results may be biased due to environmental contamination of the milk samples. Considering that the study was performed on a research farm, strict hygienic measures during the feeding experiment may have contributed to the low transfer of bacteria from feed to milk.
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The assessment of human liver stem cells (HLSCs) as cell therapeutics requires scalable, controlled expansion processes. We first focused on defining appropriate process parameters for HLSC expansion such as seeding density, use of antibiotics, optimal cell age and critical metabolite concentrations in conventional 2D culture systems. For scale-up, we transferred HLSC expansion to multi-plate and stirred-tank bioreactor systems to determine their limitations. A seeding density of 4000 cells cm-2 was needed for efficient expansion. Although growth was not significantly affected by antibiotics, the concentrations of lactate and ammonia were important. A maximum expansion capacity of at least 20 cumulative population doublings (cPDs) was observed, confirming HLSC growth, identity and functionality. For the expansion of HLSCs in the multi-plate bioreactor system Xpansion (XPN), the oxygen supply strategy was optimized due to a low kLa of 0.076 h-1. The XPN bioreactor yielded a final mean cell density of 94 ± 8 × 103 cells cm-2, more than double that of the standard process in T-flasks. However, in the larger XPN50 device, HLSC density reached only 28 ± 0.9 × 103 cells cm-2, while the glucose consumption rate increased 8-fold. In a fully-controlled 2 L stirred-tank bioreactor (STR), HLSCs expanded at a comparable rate to the T-flask and XPN50 processes in a homogeneous microenvironment using advanced process analytical technology. Ultimately, the scale-up of HLSCs was successful using two different bioreactor systems, resulting in sufficient numbers of viable, functional and undifferentiated HLSCs for therapeutic applications.
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Cell and gene therapies are an innovative solution to various severe diseases and unfulfilled needs. Adoptive cell therapy (ACT), a form of cellular immunotherapies, has been favored in recent years due to the approval of chimeric antigen receptor CAR-T products. Market research indicates that the industry's value is predicted to reach USD 24.4 billion by 2030, with a compound annual growth rate (CAGR) of 21.5%. More importantly, ACT is recognized as the hope and future of effective, personalized cancer treatment for healthcare practitioners and patients worldwide. The significant global momentum of this therapeutic approach underscores the urgent need to establish it as a practical and standardized method. It is essential to understand how cell culture conditions affect the expansion and differentiation of T-cells. However, there are ongoing challenges in ensuring the robustness and reproducibility of the manufacturing process. The current study evaluated various adoptive T-cell culture platforms to achieve large-scale production of several billion cells and high-quality cellular output with minimal cell death. It examined factors such as bioreactor parameters, media, supplements and stimulation. This research addresses the fundamental challenges of scalability and reproducibility in manufacturing, which are essential for making adoptive T-cell therapy an accessible and powerful new class of cancer therapeutics.
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This paper reviews the utility of zebrafish (Danio rerio) as a model system for exploring neurobehavioral phenomena in preclinical research, focusing on physiological processes, disorders, and neurotoxicity biomarkers. A comprehensive review of the current literature was conducted to summarize the various behavioral characteristics of zebrafish. The study examined the etiological agents used to induce neurotoxicity and the biomarkers involved, including Aß42, tau, MMP-13, MAO, NF-Ðß, and GFAP. Additionally, the different zebrafish study models and their responses to neurobehavioral analysis were discussed. The review identified several key biomarkers of neurotoxicity in zebrafish, each impacting different aspects of neurogenesis, inflammation, and neurodegeneration. Aß42 was found to alter neuronal growth and stem cell function. Tau's interaction with tubulin affected microtubule stability and led to tauopathies under pathological conditions. MMP-13 was linked to oxidative assault and sensory neuron degeneration. MAO plays a role in neurotransmitter metabolism and neurotoxicity conversion. NF-Ðß was involved in pro-inflammatory pathways, and GFAP was indicative of neuroinflammation and astroglial activation. Zebrafish provide a valuable model for neurobehavioral research, adhering to the "3Rs" philosophy. Their neurotoxicity biomarkers offer insights into the mechanisms of neurogenesis, inflammation, and neurodegeneration. This model system aids in evaluating physiological and pathological conditions, enhancing our understanding of neurobehavioral phenomena and potential therapeutic interventions.
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This study aims to assess the musculoskeletal risk of military personnel on a Leopard 2 A6 main battle tank crew and to identify associated factors for future prevention and mitigation strategies. A sample of 57 Portuguese military personnel, who are or were part of the Leopard 2 A6 main battle tank crew, answered a questionnaire on their perception of task performance, considering muscle demands, comfort, posture, movements, and associated symptoms. A subsample of four soldiers from the Armoured Squadron of the Portuguese Mechanized Brigade were assessed using an inertial measurement unit system and underwent a whole-body kinematic analysis coupled with a Rapid Entire Body Assessment during a simulated two-hour mission. The results indicate that soldiers accurately perceive their roles within the crew and that, overall, there is a high risk of musculoskeletal injuries in all tasks. However, tasks directly related to the crew's primary duties carry consistently high risk when considering the time spent on their tasks. This study highlights the need for targeted preventive measures to reduce the incidence and severity of injuries among the crew of the Leopard 2 A6 main battle tank.
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Militares , Movimento , Humanos , Fatores de Risco , Projetos Piloto , Masculino , Adulto , Movimento/fisiologia , Fenômenos Biomecânicos , Doenças Musculoesqueléticas/fisiopatologia , Doenças Musculoesqueléticas/epidemiologia , Feminino , Adulto Jovem , Inquéritos e Questionários , Análise e Desempenho de TarefasRESUMO
Herpes simplex virus type 1 (HSV-1) plays an important role in the field of gene therapy and viral vaccines, especially as an oncolytic virus. However, the mass production of HSV-1 viral vectors remains a challenge in the industry. In this study, a microcarrier-mediated serum-reduced medium culture was used to improve the bioprocess of HSV-1 production and increase HSV-1 yields. The composition of the culture media, which included a basal medium, serum concentration, and glutamine additive, was optimized. The process was successfully conducted in a 1 L bioreactor, and virus production was threefold greater than that of conventional processes with a 10% serum medium. The bead-to-bead transfer process was also developed to further increase scalability. In spinner flasks, the detachment rate increased from 49.4 to 80.6% when combined agitation was performed during digestion; the overall recovery proportion increased from 37.9 to 71.1% after the operational steps were optimized. Specifically, microcarrier loss was reduced during aspiration and transfer, and microcarriers and detached cells were separated with filters. Comparable cell growth was achieved with the baseline process using 2D culture as the inoculum by exchanging the subculture medium. To increase virus production after bead-to-bead transfer, critical parameters, including shear stress during digestion, TrypLE and EDTA concentrations in the subculture, and the CCI, were identified from 47 parameters via correlation analysis and principal component analysis. The optimized bead-to-bead transfer process achieved an average of 90.4% overall recovery and comparable virus production compared to that of the baseline process. This study is the first to report the optimization of HSV-1 production in Vero cells cultured on microcarriers in serum-reduced medium after bead-to-bead transfer. KEY POINTS: ⢠An HSV-1 production process was developed that involves culturing in serum-reduced medium, and this process achieved threefold greater virus production than that of traditional processes. ⢠An indirect bead-to-bead transfer process was developed with over 90% recovery yield in bioreactors. ⢠HSV-1 production after bead-to-bead transfer was optimized and was comparable to that achieved with 2D culture as inoculum.