Bottom Compartment Research Articles

Comparison of plastic pollution between waters and sediments in four Po River tributaries (Northern Italy)

The monitoring of plastic contamination in freshwaters is still pioneering in comparison with marine environments, and few studies analyzed the distribution of these pollutants in both aqueous and bottom compartments of continental waters. Therefore, the aim of this study was the comparison of plastic pollution in both waters and sediments of four Po River tributaries (Ticino, Adda, Oglio and Mincio Rivers), which outflow from the main Italian sub-alpine Lakes, in order to establish the strengths and weaknesses of both matrices. The main results pointed out a heterogeneous plastic contamination, with the lowest values in Ticino (0.9 ± 0.5 plastics/m3 in waters and 6.8 ± 4.5 plastics/kg dry weight - d.w. - in sediments) and the highest in Mincio (62.9 ± 53.9 plastics/m3 in waters and 26.5 ± 13.3 plastics/kg d.w in sediments), highlighting a plastic amount in sediments four times higher than waters. Plastic pollution, mainly due to microplastics, was associated principally to a domestic input in both waters and sediments of Ticino and Adda Rivers, as well as in sediments of Oglio, while an industrial pollution was found in waters and sediments of Mincio and Oglio waters. Our data clearly highlighted as the monitoring of both matrices provide complementary information for a holistic risk assessment of these emerging contaminants in freshwaters: the aqueous matrix provides an instantaneous picture of contamination, while sediments the history of pollution.

Availability and effects of n-TiO2 and PCB77 in fish in vitro models of the intestinal barrier and liver under single- and/or co-exposure scenarios

Titanium dioxide nanoparticles (n-TiO2) and polychlorinated biphenyls (PCBs) can be present in the food of fish, leading to intestinal exposure uptake, and accumulation in inner organs. This study examined combination effects of n-TiO2 and PCB77 in vitro models of the fish intestinal epithelium and liver, i.e., RTgut-GC cell cultures grown in ThinCerts™ and RTL-W1 cell cultures grown in standard tissue culture plates. Mass spectrometry and microscopy techniques were used to obtain information on nanoparticle translocation across the intestinal barrier model. In addition, the substances’ effect on intestinal barrier permeability, cell viability, expression of dioxin – and antioxidant response element -controlled genes, and induction of cytochrome P450 1a (Cyp1a)-dependent ethoxyresorufin-O-deethylase (EROD) activity were assessed. TiO2 nanoparticles were taken up by RTgut-GC cells and detected in the bottom compartment of the intestinal epithelial barrier model. It was not possible to conclude definitively if n-TiO2 translocation occurred via transcytosis or paracellular migration but observations of nanoparticles in the lateral space between adjacent epithelial cells were rare. PCB77 (1 and 10 µM, 24 h) did not affect barrier permeability, i.e., n-TiO2 translocation is probably not facilitated in case of co-exposure. Furthermore, previous and simultaneous exposure to n-TiO2 (1 and 10 mg/L, 24 h) did not have any influence on PCB77-induced Cyp1a mRNA and enzyme activity levels in RTL-W1 cells. Furthermore, there were no significant differences in expression of antioxidant response element-controlled genes comparing control, single substance, and mixture treatments, not even following long-term exposure (0.01–1 mg/L n-TiO2 + 1 nM PCB77, 4 weeks). While an underestimation of the effects of n-TiO2 and PCB77 cannot be fully excluded as concentration losses due to sorption to cell culture plastics were not measured, the results suggest that the test substances probably have a low potential to exhibit combination effects on the assessed endpoints when co-existing in fish tissues.

Treatment of petroleum refinery wastewater by adsorption using activated carbon fixed bed column with batch recirculation mode

Water pollution and the lack of access to clean water are general global problems that result from the expansion of industrial and agricultural activities. Petroleum refinery wastewaters consider as a major challenge to the environment and their treatment is mandatory. The present work concerns with the removal of chemical oxygen demand (COD) from petroleum refinery wastewater taken from Iraq's Al-Diwaniyah petroleum refinery plant by using an activated carb fixed-bed column operated at a batch recirculation mode. The fixed bed column used in this work was composed of three sections: upper, central, and bottom compartments. The bottom compartment serves as a feeding chamber to the central adsorption chamber while the upper compartment serves as a collecting effluent chamber. By adopting response surface methodology (RSM), in the pacts of various operational parameters such as packing level, pH, and time on the COD removal efficiency were investigated. The optimal conditions were an activated carbon packing level of 80%, pH of 5.7, and adsorption time of 73 min approximately, which resulted in a COD removal efficiency of 96.70%. The results indicated that the packing level of activated carbon had a major effect on COD elimination followed by pH, while time had a minor effect. The model equation's adequacy was demonstrated by its strong R2 value (0.975). The present study demonstrates that the adsorption system by activated carbon is an effective method for removingcondomODom Al-Diwaniyah petroleum refinery wastewaters.

(Digital Presentation) Effects of Thermal Convection on Species Transport in Photocatalytic Suspension Reactors

Z-scheme photocatalytic suspension reactors with aqueous redox shuttles present a promising pathway for safe and low cost solar water splitting to produce hydrogen. Soluble redox shuttles act as electron relays between light absorbing particles present in the two compartments. In recent work we have demonstrated a theoretical pathway to attain up to 4% solar-to-hydrogen efficiency with passive diffusion driven species transport [1]. However, to attain larger solar-to-hydrogen efficiencies it is crucial to enhance the rate of species transport between the two reaction compartments. We report on computational and experimental investigations to explore the effects of natural convection on species transport in Z-scheme photocatalytic systems. A transient, two-dimensional fluid flow, heat- and mass-transfer model was developed for a two-compartment system with a porous separator; horizontal and vertical reactor arrangements were considered. Volumetric absorption of visible and infrared components of incident sunlight by the suspension was modeled. Transient species concentration profiles were obtained as a function of the operating solar-to-hydrogen efficiencies, porous separator morphology and the size of the compartments. Model results predict enhanced rates of transport of redox shuttle species between two compartments in the presence of thermal convection in addition to diffusion. At 10% solar-to-hydrogen efficiency, minimum redox shuttle concentration with convection decreases by 80% as compared to passive diffusion alone for a vertically stacked reaction compartment with 2 cm height. Experimental measurements have been also performed on two-compartment prototypes (horizontal and vertical) to probe the effects of natural convection on species mixing. The source of illumination was a xenon-lamp solar simulator (Newport 94023A) with an AM1.5G filter and calibrated such that the incident illumination power at the top of the reactor was 1 sun (1000 W/m2). The reactor was assembled by connecting two polycarbonate tubes (5 cm x 5 cm) with a height of 5 cm and placing a thin separator (Nafion 117 and regenerated cellulose) between the tubes to separate the compartments. A base was 3D-printed and painted black to secure the bottom compartment and maximize absorption of the incident illumination and promote natural convection. Thermocouples (K-type) were placed at various heights within the two compartments to track local temperatures within the fluid, including along the bottom surface of the lower compartment. Over a 3-hour period of 1 sun irradiance, the temperatures were recorded, and a favorable temperature gradient for natural convection was observed (ΔT ~ 2 °C) for vertically-stacked reaction compartments. The flow behavior was visualized through the injection of a colored dye at the bottom surface of the reactor and above the membrane after the 3-hour period. Image processing of the convection video is used to help quantify the rate of mixing within the compartments. Model predictions for flow behavior and temperature profiles were compared to and validated against experimental measurements. Collectively, this study implements a unique approach to predict and evaluate the effect of natural convection on species transport in Z-scheme photocatalytic reactors.[1] Bala Chandran, R., Breen, S., Shao, Y., Ardo, S., & Weber, A. Z. (2018). Evaluating particle-suspension reactor designs for Z-scheme solar water splitting via transport and kinetic modeling. Energy and Environmental Science, 11(1), 115–135. https://doi.org/10.1039/c7ee01360d Figure 1

Combining Human Organoids and Organ-on-a-Chip Technology to Model Intestinal Region-Specific Functionality

The intestinal mucosa is a complex physical and biochemical barrier that fulfills a myriad of important functions. It enables the transport, absorption, and metabolism of nutrients and xenobiotics while facilitating a symbiotic relationship with microbiota and restricting the invasion of microorganisms. Functional interaction between various cell types and their physical and biochemical environment is vital to establish and maintain intestinal tissue homeostasis. Modeling these complex interactions and integrated intestinal physiology in vitro is a formidable goal with the potential to transform the way new therapeutic targets and drug candidates are discovered and developed. Organoids and Organ-on-a-Chip technologies have recently been combined to generate human-relevant intestine chips suitable for studying the functional aspects of intestinal physiology and pathophysiology in vitro. Organoids derived from the biopsies of the small (duodenum) and large intestine are seeded into the top compartment of an organ chip and then successfully expandas monolayers while preserving the distinct cellular, molecular, and functional features of each intestinal region. Human intestine tissue-specific microvascular endothelial cells are incorporated in the bottom compartment of the organ chip to recreate the epithelial-endothelial interface. This novel platform facilitates luminal exposure to nutrients, drugs, and microorganisms, enabling studies of intestinal transport, permeability, and host-microbe interactions. Here, a detailed protocol is provided for the establishment of intestine chips representing the human duodenum (duodenum chip) and colon (colon chip), and their subsequent culture under continuous flow and peristalsis-like deformations. We demonstrate methods for assessing drug metabolism and CYP3A4 induction in duodenum chip using prototypical inducers and substrates. Lastly, we provide a step-by-step procedurefor the in vitro modeling of interferon gamma (IFNγ)-mediated barrier disruption (leaky gut syndrome) in a colon chip, including methods for evaluating the alteration of paracellular permeability, changes in cytokine secretion, and transcriptomic profiling of the cells within the chip.

Acanthamoeba castellanii: Effect of neuroactive substances on trophozoite migration

Acanthamoeba castellanii is the etiological agent of granulomatous amebic encephalitis, amebic keratitis, and skin lesions. In vitro and in vivo studies have demonstrated that Acanthamoeba trophozoites induce contact-dependent, and contact-independent pathogenic mechanisms. We have explored the potential role neuroactive substances may have in the migration of Acanthamoeba castellanii trophozoites using Transwell permeable supports in the presence of physiological concentrations of dopamine, glutamate, serotonin, or taurine diluted in PBS. Quantitation of migrated amoebae was carried out in scanning electron micrographs of the upper and under compartments sides of the chamber membranes. Our results showed that at 2 h of interaction, a statistically significant larger proportion of A. castellanii trophozoites migrated through the chamber membranes when neurotransmitters were placed in the lower compartments of the chambers compared to control. This migration effect was more evident under the presence of glutamate and taurine on the three surfaces (upper/lower membrane and bottom compartment) when the percentage of migrated trophozoites was analyzed. Scanning electron microscopy of trophozoites revealed that glutamate and taurine induced the formation of large adhesion lamellas and phagocytic stomas. These observations suggest that certain neuroactive substances could stimulate the migration of A. castellanii trophozoites in the central nervous system.

Transfer of La, Ce, Sm and Yb to alfalfa and ryegrass from spiked soil and the role of Funneliformis mosseae.

Rare earth elements (REEs) are widely used in high-tech industries, and REE waste emissions have become a concern for ecosystems, food quality and human beings. Arbuscular mycorrhizal fungi (AMF) have repeatedly been reported to alleviate plant stress in metal-contaminated soils. To date, little information is available concerning the role of AMF in REE-contaminated soils. We recently showed that there was no transfer of Sm to alfalfa by Funneliformis mosseae, but only a single REE was examined, while light and heavy REEs are present in contaminated soils. To understand the role of AMF on the transfer of REEs to plants, we carried out an experiment using alfalfa (Medicago sativa) and ryegrass (Lolium perenne) in compartmented pots with separate bottom compartments that only were accessible by F. mosseae fungal hyphae. The bottom compartments contained a mixture of four REEs at equal concentrations (La, Ce, Sm and Yb). The concentration of REEs in plants was higher in roots than in shoots with higher REE soil-root than root-shoot transfer factors. Moreover, significantly higher light-REEs La and Ce were transferred to ryegrass shoots than Sm and the heavy-REE Yb, but this was not observed for alfalfa. Alfalfa dry weight was significantly increased by F. mosseae inoculation, but not ryegrass dry weight. For both plant species, there was significantly higher P uptake by the mycorrhizal plants than the nonmycorrhizal plants, but there was no significant transfer of La, Ce, Sm or Yb to alfalfa and ryegrass roots or shoots due to F. mosseae inoculation.

The hydraulic lift response under media moisture conditions on the cashew (Anacardium occidentale L.)

Cashew was reported to have hydraulic lift (HL) ability, which vertically transferred water from the moister bottom layer to the drier upper one. However, it was tested under homogenous moisture conditions on the bottom soil layer, whereas its response under heterogeneous one was still unclear. Therefore, this study evaluated the HL response under heterogeneous moisture on the bottom soil layer on 10 months old cashew. It was conducted in the ISMCRI greenhouse from September to October 2020. The cashew was grown, and its roots were developed into three compartments filled with growth media: one top compartment (T) and two bottom compartments (B1 and B2). The media was subjected to four conditions 1) the whole media in moist conditions, 2) T was dry, B1 and B2 were moist, 3) T and B1 were dry, B2 was moist, and 4) the whole media were dry. The cashew HL was not significantly reduced in the condition where the half part of the lower root was in dry condition. This result indicated that cashew HL was strong enough under heterogeneous soil moisture conditions in the bottom layer. Thus, cashew was potential as a component of water bio-conservation of the land in the dry climate.

‘Stirred not Shaken!’ Comparing Agitation Methods for Permeability Studies Using a Novel Type of 96-Well Sandwich-Plates

In order to achieve a high sample throughput, permeation experiments are often carried out using 96-well sandwich plates. Even though agitation is regarded as important, permeation studies in 96-well format are often carried out without agitation since orbital shaking, the most common agitation method for 96-well plates, has been reported to create difficulties (e.g., well-to-well cross-talk), and high cost and low availability limits the use of other agitation techniques (e.g., magnetic stirring). This study investigates how orbital shaking and magnetic stirring affect the apparent permeability of model compounds with different water-solubilities (methylene blue, carbamazepine, and albendazole) using a novel 96-well sandwich plate comprising a cellulose-hydrate membrane (PermeaPlain® plate). Orbital shaking was found less efficient than magnetic stirring in terms of homogeneously distributing a small volume of dye within the donor compartment. Furthermore, in terms of achieving maximum trans-barrier flux, magnetic stirring was found a more effective agitation method than orbital shaking. Obviously, with orbital shaking the medium in the bottom compartment of the sandwich plates never was mixed in-phase. The impact of insufficient mixing on permeation was found strongest with the most lipophilic compound, which correlates with literature reports that the contribution of the unstirred water layer towards the overall resistance of the barrier is most expressed in case of lipophilic drugs. Finally, it was tested how different liquid volumes in the bottom compartment of the plates affect the well-to-well cross-talk during permeation experiments under orbital shaking. This study revealed that 250–300 µL should be used in the bottom compartment of the sandwich plates to reduce well-to-well cross-talk when using orbital shaking for agitation.

Rapid Truck Loading for Efficient Feedstock Logistics

A multi-bale handling unit offers an advantage for the efficient hauling of round bales. Two empty racks on trailers are left at a satellite storage location for loading while a truck tractor delivers two loaded racks to the biorefinery, thus uncoupling the loading and hauling operations and increasing the efficiency of both. The projected 10 min trailer exchange time equals the projected 10 min unload time at the biorefinery achieved by lifting off the two full racks and replacing them with two empties, a technology adapted from the container shipping industry. A concept is presented for a bale loader that latches onto the rack/trailer and loads bales into the bottom tier chambers. This machine will load 10 bales into the rack on the front trailer by attaching on to the front of the trailer and 10 bales into the rear trailer by attaching onto the rear. A telehandler removes bales from single-layer storage and places them in the bale loader to load the bottom tier compartments. The top tier compartments are loaded directly from the top. Expectations are that an experienced operator can average 9 loads in a 10 h workday, and load-out cost is estimated as 3.61 USD/Mg, assuming the average achieved load-out productivity over annual operation is 60% of optimum productivity (24 Mg/h) equal to 14.4 Mg/h. Cost increases to 4.81 USD/Mg when the productivity factor drops to 45%, and cost is 3.09 USD/Mg for a factor of 70%.

Real-time quantitative monitoring of in vitro nasal drug delivery by a nasal epithelial mucosa-on-a-chip model

ABSTRACT Objectives A human nasal epithelial mucosa (NEM) on-a-chip is developed integrated with a novel carbon nanofibers-modified carbon electrode for real-time quantitative monitoring of in vitro nasal drug delivery. The integration of platinum electrodes in the chip also enables real-time measurement of transepithelial electrical resistance (TEER). Methods The air-liquid interface culture of nasal epithelial RPMI 2650 cells in the NEM-on-a-chip was optimized to mimic the key functional characteristics of the human nasal mucosa. The epithelial transport of ibuprofen in the NEM-on-a-chip was electrochemically monitored in real-time under static and physiologically realistic dynamic flow conditions. Results The NEM-on-a-chip mimics the mucus production and nasal epithelial barrier function of the human nasal mucosa. The real-time drug quantification by the NEM-on-a-chip was validated versus the high-performance liquid chromatography method. The drug transport rate monitored in the NEM-on-a-chip was influenced by the flow in the bottom compartment of the chip, highlighting the importance of emulating the dynamic in vivo condition for nasal drug transport studies. Conclusion This novel NEM-on-a-chip can be a low-cost and time-efficient alternative to the costly laborious conventional techniques for in vitro nasal drug transport assays. Importantly, its dynamic microenvironment enables conducting nasal drug transport tests under physiologically relevant dynamic conditions.

A novel approach based on multiple fish species and water column compartments in assessing vertical microlitter distribution and composition.

The assessment of the distribution and composition of microlitter in the sea is a great challenge. Biological indicators can be an irreplaceable tool since they measure microlitter levels in their environments in a way that is virtually impossible to replicate by direct physical measurements. Furthermore, trends can provide policymakers with statistically robust analysis. We looked into the capacity of multiple fish species to describe the distribution and composition of microlitter vertically across different compartments of the water column. A total of 502 individuals from six selected species (Scomber scombrus, Oblada melanura, Spicara smaris, Boops boops, Merluccius merluccius and Mullus barbatus) were collected on the western side of Sardinia island and allocated to three compartments: surface, mid-water and bottom. The species of the surface exhibited a higher frequency of occurrence (41.89%) of microlitter ingestion, compared to those of the mid-water and bottom (19.60%; 22.58%). A significant difference in the average number of ingested microlitter was found between the surface and the bottom compartment. All the microlitter fragments found were analysed through Fourier Transform Infrared Spectroscopy (FTIR). The comparison of the expected buoyancies of the polymers identified puth faith in the allocation of the species to the respective compartments. Therefore, considering the Marine Strategy Framework Directive objective, this approach could be useful in assessing microlitter distribution and composition vertically across the water column.

Study of the vertical temperature profile in a tomato greenhouse equipped with lighting, two screens and a VentilationJet system

The application of Next Generation Growing (NGG, in Dutch “Het Nieuwe Telen”) in Dutch greenhouses has resulted in energy savings, accompanied by the extensive use of screens. The latter has affected the control of greenhouse climate which strongly depends on the air and humidity exchange between the top and bottom greenhouse compartment, as they are separated by the screen itself. When the screens are fully deployed an air exchange/mix system (VentilationJet) that blows dry and cold air from the top compartment into the greenhouse can be used to lower the greenhouse air temperature and relative humidity in a controllable way. The effect of this greenhouse climate control equipment use on vertical air temperature profile as well as on energy use was studied in a commercial greenhouse. Effects of artificial lighting, heating with pipes below the crop, activation of vertical fans and air exchange rate were analyzed during the winter of 2018. It was observed that when significant heat input (radiation) at the top of the crop occurred a vertical temperature gradient up to 2°C exists, with the bottom of the crop being colder than the top. This temperature gradient cannot be reduced by using only vertical air circulation fans but it can be minimized by additional heat input at the bottom; this may result in excess heat that has to be removed through either natural ventilation or with the use of VentilationJet when the screens are deployed. During the studied period about a quarter of the daily gas use for heating took place at the same time as the VentilationJet was removing warm and humid air from the top of the greenhouse.

P006 3D Multicellular intestine-on-a-chip model for disease modelling and drug discovery

Abstract Background One of the major functions of the human intestine is to provide a protective epithelial barrier between the body and digestive environment. Additionally, the interplay of commensal microbes of the gut microbiome with the gut tissue and host immune system significantly contributes to intestinal homeostasis. Crohn’s disease and ulcerative colitis, collectively referred to as inflammatory bowel diseases, are both associated with increased permeability of the epithelial barrier and dysregulated immune response. Great efforts have been made to develop both in vitro and in vivo models of the human intestine in order to understand the development and underlying pathogenesis of IBD. These efforts have provided valuable insights into multiple aspects of the disease. However, none of these models has been able to capture the complexity and multifactorial nature of IBD. Animal models generally fail to accurately predict the efficacy and toxicity of novel compounds in human tissues, while in vitro human intestinal models developed on porous membranes within Transwell inserts fail to accurately recapitulate and mimic key aspects of the in vivo situation. Methods Here, we present the development and characterisation of a 3D multicellular perfused intestine-on-a-chip model in a microfluidic platform, the OrganoPlate®, and its application for investigating intestinal inflammation. The model described here comprises a coculture of Caco-2 and mucus-secreting HT29-MTX cells in the top compartment of the chip and a coculture of immune-competent cells THP-1 and MUTZ-3 in the bottom compartment, lining a collagen-I ECM in the middle. Results We show that the Caco-2 and HT29-MTX coculture form confluent and polarised tubular structures against the collagen-I ECM in the OrganoPlate®, with a stable barrier function over time as well as the capability to secrete mucus. By exposing the cultures to TNFα and/or IL-1β, we were able to induce an inflammatory state, characterised by cytokine release (IL-8) and a decrease in trans-epithelial electrical resistance. Finally, we proved the applicability of the model in screening anti-inflammatory compounds by its reversibility. Using a well-known anti-inflammatory drug, TPCA-1, we were able to prevent cytokine-induced inflammation. This result was evident from the decreased secretion of IL-8 and retention of barrier function in treated cultures, similar to that observed in untreated cultures. Conclusion Overall, this complex 3D multicellular perfused intestine-on-a-chip model provides the versatile modularity of mimicking key features of intestinal inflammation and can, therefore, further support drug screening efforts and provide a platform for personalised medicine.

Quantification of hydraulic redistribution in maize roots using neutron radiography

AbstractPlants redistribute water from wet to dry soil layers through their roots, in the process called hydraulic redistribution. Although the relevance and occurrence of this process are well accepted, resolving the spatial distribution of hydraulic redistribution remains challenging. Here, we show how to use neutron radiography to quantify the rate of water efflux from the roots to the soil. Maize (Zea mays L.) plants were grown in a sandy substrate 40 cm deep. Deuterated water (D2O) was injected in the bottom wet compartment, and its transport through the roots to the top dry soil was imaged using neutron radiography. A diffusion–convection model was used to simulate the transport of D2O in soil and root and inversely estimate the convective fluxes. Overnight, D2O appeared in nodal and lateral roots in the top compartment. By inverse modeling, we estimated an efflux from lateral roots into the dry soil equal to jr = 2.35 × 10−7 cm−1. A significant fraction of the redistributed water flew toward the tips of nodal roots (3.85 × 10−8 cm3 s−1 per root) to sustain their growth. The efflux from nodal roots depended on the roots’ length and growth rate. In summary, neutron imaging was successfully used to quantify hydraulic redistribution. A numerical model was needed to differentiate the effects of diffusion and convection. The highly resolved images showed the spatial heterogeneity of hydraulic redistribution.

Developing a method of co‐incubating murine adipocytes and macrophages for evaluating cellular communication

Adipose tissue, an organ containing both adipocytes and adipose tissue macrophages (ATMs), secretes cytokines known as adipokines. Of these two cell types, ATMs have a larger role in the production and release of proinflammatory adipokines. The objective of this research was to develop a system of co‐incubating adipocytes and macrophages that allowed us to quantitatively measure the cross‐talk between these two cell types. Nitric Oxide (NO) and interleukin‐6 (IL‐6) concentrations were quantified to determine macrophage activation and production of a proinflammatory adipokine, respectively. Leptin concentrations were used as a measure of adipocyte‐specific adipokine production. The cell lines used were RAW 264.7 (murine macrophages) and 3T3‐L1 (murine fibroblasts). 3T3‐L1 fibroblasts were differentiated into adipocytes, which was confirmed by the presence of lipid droplets. Transwell plates with permeable inserts were used to co‐incubate the macrophages and adipocytes; the confluent adipocytes were maintained in the 12‐well plate (lower compartment) and the macrophages were added to the transwell insert (upper compartment). Macrophages were stimulated with 0.01 mg/ml of LPS either before or after the addition of the the macrophages to the insert. The treatments were: 3T3+media, 3T3+media+LPS, 3T3+unstimulatedRAW, 3T3+RAW+LPS, 3T3+stimulatedRAW (in fresh media), and 3T3+stimulatedRAW (in conditioned media). There was no difference in IL‐6 concentration measured in the upper compartment of the transwell insert vs the lower compartment, while measured NO levels were higher in the upper compartment and leptin levels were higher in the lower compartment (p<0.001, matched pairs t‐test). The 3T3+RAW+LPS, 3T3+stimulatedRAW (in fresh media) and 3T3+stimulatedRAW (in conditioned media) co‐incubation protocols resulted in the highest concentration of IL‐6 (p=0.0006). The 3T3+RAW+LPS and 3T3+stimulatedRAW (in conditioned media) co‐incubation methods yielded the highest concentrations of NO in both the upper and bottom compartments of the plate (p<0.001). There was no difference in leptin concentrations due to co‐incubation protocol. There was a positive relationship between NO and IL‐6 in both the top and bottom compartments (p<0.001, linear regression) and between NO and leptin in the top (p=0.03) but not in the bottom compartment. There was no significant relationship between IL‐6 and leptin concentrations in either the top or bottom compartments. For future experiments, our lab will utilize the co‐incubation protocols that resulted in the highest production of NO and IL‐6: 3T3+RAW+LPS and 3T3+stimulatedRAW (in conditioned media).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Indirect effect of PM1 on endothelial cells via inducing the release of respiratory inflammatory cytokines

A large number of epidemiological studies have shown that increased cardiovascular morbidity and mortality are associated with exposure to high concentrations of PM2.5. One of the ways that PM2.5 affects the cardiovascular system is through systemic inflammation. Inflammatory cytokines such as TNF-α, IL-1β, IL-6, and IL-8 stimulate endothelial cells, which leads to endothelial dysfunction. Compared with PM2.5, PM1 is smaller in size, has a larger surface area and absorbs more toxic substances such as heavy metals, organic compounds, and black carbon. However, the effect of PM1 on human health is less studied. Here, we used BEAS-2B cells and differentiated THP-1 cells to simulate epithelial cells and macrophages in the lung, respectively. The indirect effect of PM1 on endothelial cells was studied with a coculture model consisting of two cell lines (BEAS-2B cells and macrophages) in the top compartment and one cell line, human umbilical vein endothelial cells (EA.hy926), in the bottom compartment of a transwell plate. The results showed that PM1 could promote the release of inflammatory cytokines, including TNF-α and IL-6, from BEAS-2B cells and macrophages. In addition, PM1 upregulated ICAM-1 expression in EA.hy926 cells through TNF-α/NF-κB signaling pathways, promoting the adhesion of endothelial cells and monocytes, a key event in the initiation of atherosclerosis.

Sensitivity of multispecies maximum sustainable yields to trends in the top (marine mammals) and bottom (primary production) compartments of the southern North Sea food-web

In marine ecosystems, maximum sustainable yield considerations are affected by any substantial changes that occur in the top and bottom compartments of the food-web. This study explores how the southern North Sea’s fisheries may need to adjust their fishing efforts to maintain optimum yields of sole, plaice, cod and brown shrimps under increased marine mammal populations and a reduced primary productivity. We constructed plausible scenarios of ongoing food-web changes using the results of Bayesian age-structured population models to estimate carrying capacities of harbour porpoises (Phocoena phocoena) and grey seals (Halichoerus grypus). Losses in primary productivity were predicted by lower trophic level ecosystem models. These scenarios were implemented in a food-web model of the southern North Sea. For each scenario, we sought mixed-fleet fishing efforts that would deliver maximum yields of sole, plaice, cod and brown shrimp combined. We also did so for a baseline run with unaltered mammal and primary production, and compared the differences in optimal fishing strategies, predicted yields, and states of the stocks between the scenarios. We found stocks and yields to be far more sensitive to changes in primary productivity than to increased marine mammal predation. The latter predominantly impacted cod, and even benefitted brown shrimps compared to the baseline run. Under 30% reduced primary productivity, fishing efforts had to be reduced by 50% to still provide maximum yields, whereas the marine mammal scenario induced no need to adjust the fishing regime. This draws attention to the potential gains of incorporating bottom-up processes into long-term management considerations, while marine mammal predation may be less of a concern, in particular for flatfish fisheries in the North Sea, and may even benefit shrimp trawlers because of reduced predation on shrimp from fish predators.

Host-Pathogen Interaction Reconstituted in Three-Dimensional Cocultures of Mucosa and Candida albicans.

Candida albicans frequently causes recurrent intimal infectious disease (ID). This demands the treatment of multiple phases of the infection. The objective of this study was to uncover the host-pathogen interaction using two-dimensional (2D) epithelium cell-barrier and three-dimensional (3D) subepithelium tissue cells of human mucosa. The 2D cell cultures assessed C. albicans adhesion. Addition of the antifungal drug Fluconazol did not inhibit the adhesion, despite its pathogen growth inhibition (minimal inhibitory concentration value 0.08 μg/mL). A 3D tissue was engineered in multitranswells by placing human fibroblast cultures on a thick porous scaffold. This contained the yeast placed in the top compartment and prevented passive penetration. After 28 h, the pathogen transmigrated the barrier and was collected in the bottom compartment. A change in pathogen morphology was observed where hypha formed and grew to be 231 μm long after 28 h. The hypha was thus long enough to cross the 200 μm thick 3D tissue. The 3D infection was inhibited by addition of Fluconazol (0.08 μg/mL), confirming that penetration is dependent on pathogen growth. In conclusion, ID was reconstituted step-by-step on 2D epithelium surface and in 3D connective tissue of human mucosa. Fluconazol growth-inhibition of the pathogen C. albicans was confirmed in the 3D tissue. We thus propose that this ID in vitro test is suitable for the identification and characterization of new treatments against C. albicans.

The POSEIDON-R compartment model for the prediction of transport and fate of radionuclides in the marine environment

A detailed description of the advanced version of compartment model POSEIDON-R for the prediction of transport and fate of radionuclides in the marine environment is given. The equations of transfer of radionuclides in the water and bottom sediment compartments along with the dynamical food chain model are presented together with dose module to assess individual and collective doses to the population due to the regular and accidental releases of radionuclides. The method for the numerical solution of model equations is also presented. The modelling results for the northeast Atlantic shelf seas were compared with measurements of 137Cs.•The three-dimensional compartment model POSEIDON-R describes the transfer of radionuclides and their daughter products in marine environment as a results of regular or accidental releases. This includes any transfer through the water column and sediments.•The model is complemented by a dynamic food chain model for transfer of radioactivity in pelagic and benthic food webs.•The dose module in the model calculates internal and external doses for humans and non-human biota.