Impacts of industrial atmospheric emissions on watershed export of dissolved ions in coastal streams: a Bayesian modeling approach.

Anthropogenic atmospheric emission and subsequent deposition of sulfur (S) has been linked to disrupted watershed biogeochemical processes through soil and surface water acidification. We investigated watershed-scale impacts of acidic deposition on tributary concentrations and watershed exports of major nutrients and ions for the Kitimat River Watershed, British Columbia. Since the 1950s, the Kitimat watershed had an aluminum smelting facility with substantial emissions at the river estuary. Emissions load the airshed overlying the watershed and potentially impact western tributaries leaving eastern tributaries available as reference. We assessed concentrations and export of key compounds in three reference and six potentially impacted tributaries and watersheds in 2015 and 2016. Sulfate (SO4), fluoride (F), nitrate (NO3), and chloride (Cl) were significantly higher in impacted tributaries. F concentrations exceeded the Canadian Council of Ministers of the Environment guideline for aquatic life in 83% of samples collected from impacted streams. Watershed export and associated uncertainty were determined by bootstrapped flow-stratified Beale's unbiased estimator. Impact of emissions on watershed export was modeled in a Bayesian approach to include variance in the export estimate to inform the uncertainty of model parameters. Export of SO4 and Ca increased significantly within 16 km and 8 km, respectively, toward the smelter emissions. The corresponding impacted area for SO4 and Ca was approximately 100 km2 and 45 km2, respectively. SO4 export is likely due to direct impacts of S deposition, with excess S being flushed from the watersheds. Ca export patterns likely result from indirect impacts of S deposition on soil chemistry and flushing of Ca. These impacts may contribute to effects within tributaries on benthic stream communities and regionally important juvenile Pacific salmon.

Low Greenhouse Gas Emissions from Oyster Aquaculture.

Production of animal protein is associated with high greenhouse gas (GHG) emissions. Globally, oyster aquaculture is increasing as a way to meet growing demands for protein, yet its associated GHG-emissions are largely unknown. We quantified oyster aquaculture GHG-emissions from the three main constituents of GHG-release associated with terrestrial livestock production: fermentation in the animal gut, manure management, and fodder production. We found that oysters release no methane (CH4) and only negligible amounts of nitrous oxide (0.00012 ± 0.00004 µmol N2O gDW-1 hr-1) and carbon dioxide (3.556 ± 0.471 µmol CO2 gDW-1 hr-1). Further, sediment fluxes of N2O and CH4 were unchanged in the presence of oyster aquaculture, regardless of the length of time it had been in place. Sediment CO2 release was slightly stimulated between 4 and 6 years of aquaculture presence and then returned to baseline levels but was not significantly different between aquaculture and a control site when all ages of culture were pooled. There is no GHG-release from oyster fodder production. Considering the main drivers of GHG-release in terrestrial livestock systems, oyster aquaculture has less than 0.5% of the GHG-cost of beef, small ruminants, pork, and poultry in terms of CO2-equivalents per kg protein, suggesting that shellfish aquaculture may provide a a low GHG alternative for future animal protein production compared to land based sources. We estimate that if 10% of the protein from beef consumption in the United States was replaced with protein from oysters, the GHG savings would be equivalent to 10.8 million fewer cars on the road.

Urban groundwater dissolved silica concentrations are elevated due to vertical composition of historic land-filling.

Human influences on global silicon (Si) cycling include land-use change, deforestation, and wastewater discharge. Here we quantified the effect of urban expansion and historic land fill on dissolved silica (DSi) concentrations in urban groundwater in a northern temperate city. We hypothesized that historical land use, fill material, and urban infrastructure buried below cities create a unique anthropogenic geology which acts as a DSi source. We found that concentrations of DSi in urban groundwater are significantly higher than those from non-urban environments. We also found that historic land-use variables out-perform traditional topographic variables predicting urban DSi concentrations. We show that higher groundwater DSi concentrations result in increased subterranean groundwater discharge (SGD) fluxes, thereby altering coastal receiving water DSi availability. Further, we demonstrate that accounting for urban SGD DSi fluxes globally, could increase DSi SGD export by 20%. Together these results call for a re-evaluation of anthropogenic impacts on the global Si cycle.

Distinguishing point and non-point sources of dissolved nutrients, metals, and legacy contaminants in the Detroit River.

Water quality impacts to the Laurentian Great Lakes create bi-national issues that have been subject of investigation since the 1970s. However, distinguishing upgradient sources of nutrients, metals and legacy contaminants in rivers remains a challenge, as they are derived from multiple sources and flows typically vary throughout the region. These complications are especially pertinent in the Lake Huron to Lake Erie corridor and Detroit River. The Detroit River supplies 90% of the water to the western basin of Lake Erie (5300 m3/s) and is subject to a variety of co-occurring potential sources (e.g., agriculture, urbanization, and upgradient water bodies) of water quality indicators that limit source disaggregation. To find the source signal in the noise we used an integrative interpretation of dissolved chemical and isotopic parameters with sediment chemical, isotopic, and contaminant indicators. The approach combines archival data to distinguish point and non-point sources, and upgradient water bodies as sources of nutrients, metals and contaminants to the Detroit River and ultimately the western basin of Lake Erie. Persistent organic pollutants and metals cluster together as an urban group. Regional dissolved orthro-phosphate (PO4) in the water column also groups with urban point sources rather than agricultural sources. Urbanization as the primary source of PO4 in the Detroit River highlights the need for continued research on urban impacts and assessments of broader best management practices protecting Lake Erie.

Increased nutrient concentrations in Lake Erie tributaries influenced by greenhouse agriculture.

Greenhouse production of vegetables is a growing global trade. While greenhouses are typically captured under regulations aimed at farmland, they may also function as a point source of effluent. In this study, the cumulative impacts greenhouse effluents have on riverine macronutrient and trace metal concentrations were examined. Water samples were collected Bi-weekly for five years from 14 rivers in agriculturally dominated watersheds in southwestern Ontario. Nine of the watersheds contained greenhouses with their boundaries. Greenhouse influenced rivers had significantly higher concentrations of macronutrients (nitrogen, phosphorus, and potassium) and trace metals (copper, molybdenum, and zinc). Concentrations within greenhouse influenced rivers appeared to decrease over the 5-year study while concentrations within non-greenhouse influenced river remained constant. The different temporal pattern between river types was attributed to increased precipitation during the study period. Increases in precipitation diluted concentrations in greenhouse influenced rivers; however, non-influenced river runoff proportionally increased nutrient mobility and flow, stabilizing the observed concentrations of non-point sources. Understanding the dynamic nature of environmental releases of point and non-point sources of nutrients and trace metals in mixed agricultural systems using riverine water chemistry is complicated by changes in climatic conditions, highlighting the need for long-term monitoring of nutrients, river flows and weather data in assessing these agricultural sectors.

Design and Evaluation of a Robotic Device for Automated Tail Vein Cannulations in Rodent Models.

Preclinical testing in rodent models is a ubiquitous part of modern biomedical research and commonly involves accessing the venous bloodstream for blood sampling and drug delivery. Manual tail vein cannulation is a time-consuming process and requires significant skill and training, particularly since improperly inserted needles can affect the experimental results and study outcomes. In this paper, we present a miniaturized, robotic medical device for automated, image-guided tail vein cannulations in rodent models. The device is composed of an actuated three degrees-of-freedom (DOFs) needle manipulator, three-dimensional (3D) near-infrared (NIR) stereo cameras, and an animal holding platform. Evaluating the system through a series of workspace simulations and free-space positioning tests, the device exhibited a sufficient work volume for the needle insertion task and submillimeter accuracy over the calibration targets. The results indicate that the device is capable of cannulating tail veins in rodent models as small as 0.3 mm in diameter, the smallest diameter vein required to target.

Fate and Effect of Dissolved Silicon within Wastewater Treatment Effluent.

In large rivers, the ratios of silicon (Si)/nitrogen (N)/phosphorus (P) have changed dramatically as anthropogenic additions of N or P are not matched by Si. Wastewater effluent is a recognized source of N and P to coastal environments. Few previous studies, however, have examined the Si load of a large wastewater plant's effluent or the molar ratios of Si/N and Si/P in effluent. We examine the annual flux of dissolved silicon (DSi) carried by effluent from the second largest treatment plant by flow in the United States (Deer Island Treatment Plant, DITP, Boston, MA). We compare treatment plant nutrient fluxes to local urban river nutrient fluxes and trace the impact of the DITP DSi loading on receiving waters. Estimates (±95% confidence interval) of treated effluent (67 800 ± 1500 kmol DSi year-1) compared to untreated (69 500 kmol DSi year-1) indicate that the process of sewage treatment at DITP likely does not remove DSi. DITP effluent was Si-limited and this Si-limitation is reflected in the receiving waters (Massachusetts Bay). However, Si-limitation appears only in the area immediately surrounding the effluent discharge. We use these results to explain phytoplankton patterns in Massachusetts Bay and to provide the first estimate of DSi loading (3.6 Gmol SiO2 year-1) from wastewater effluent across the US.

CFD assessment of the effect of convective mass transport on the intracellular clearance of intracellular triglycerides in macrosteatotic hepatocytes.

Donor livers available to transplant for patients with end-stage liver disease are in severe shortage. One possible avenue to expand the donor pool is to recondition livers that would be otherwise discarded due to excessive fat content. Severely steatotic livers (also known as fatty livers) are highly susceptible to ischemia-reperfusion injury and as a result, primary liver non-function post-transplantation. Prior studies in isolated perfused rat livers suggest that "defatting" may be possible in a timeframe of a few hours; thus, it is conceivable that fatty liver grafts could be recovered by machine perfusion to clear stored fat from the organ prior to transplantation. However, studies using hepatoma cells and adult hepatocytes made fatty in culture report that defatting may take several days. Because cell culture studies were done in static conditions, we hypothesized that the defatting kinetics are highly sensitive to flow-mediated transport of metabolites. To investigate this question, we experimentally evaluated the effect of increasing flow rate on the defatting kinetics of cultured HepG2 cells and developed an in silico combined reaction-transport model to identify possible rate-limiting steps in the defatting process. We found that in cultured fatty HepG2 cells, the time required to clear stored fat down to lean control cells can be reduced from 48 to 4-6 h by switching from static to flow conditions. The flow required resulted in a fluid shear of .008 Pa, which did not adversely affect hepatic function. The reaction-transport model suggests that the transport of L-carnitine, which is the carrier responsible for taking free fatty acids into the mitochondria, is the key rate-limiting process in defatting that was modulated by flow. Therefore, we can ensure higher levels of L-carnitine uptake by the cells by choosing flow rates that minimize the limiting mass transport while minimizing shear stress.

Adaptive Kinematic Control of a Robotic Venipuncture Device Based on Stereo Vision, Ultrasound, and Force Guidance.

Robotic systems have slowly entered the realm of modern medicine; however, outside the operating room, medical robotics has yet to be translated to more routine interventions such as blood sampling or intravenous fluid delivery. In this paper, we present a medical robot that safely and rapidly cannulates peripheral blood vessels-a procedure commonly known as venipuncture. The device uses near-infrared and ultrasound imaging to scan and select suitable injection sites, and a 9-DOF robot to insert the needle into the center of the vessel based on image and force guidance. We first present the system design and visual servoing scheme of the latest generation robot, and then evaluate the performance of the device through workspace simulations and free-space positioning tests. Finally, we perform a series of motion tracking experiments using stereo vision, ultrasound, and force sensing to guide the position and orientation of the needle tip. Positioning experiments indicate sub-millimeter accuracy and repeatability over the operating workspace of the system, while tracking studies demonstrate real-time needle servoing in response to moving targets. Lastly, robotic phantom cannulations demonstrate the use of multiple system states to confirm that the needle has reached the center of the vessel.

3D Near Infrared and Ultrasound Imaging of Peripheral Blood Vessels for Real-Time Localization and Needle Guidance.

This paper presents a portable imaging device designed to detect peripheral blood vessels for cannula insertion that are otherwise difficult to visualize beneath the skin. The device combines near infrared stereo vision, ultrasound, and real-time image analysis to map the 3D structure of subcutaneous vessels. We show that the device can identify adult forearm vessels and be used to guide manual insertions in tissue phantoms with increased first-stick accuracy compared to unassisted cannulation. We also demonstrate that the system may be coupled with a robotic manipulator to perform automated, image-guided venipuncture.

Multilayered tissue mimicking skin and vessel phantoms with tunable mechanical, optical, and acoustic properties.

PURPOSE: This paper describes the design, fabrication, and characterization of multilayered tissue mimicking skin and vessel phantoms with tunable mechanical, optical, and acoustic properties. The phantoms comprise epidermis, dermis, and hypodermis skin layers, blood vessels, and blood mimicking fluid. Each tissue component may be individually tailored to a range of physiological and demographic conditions. METHODS: The skin layers were constructed from varying concentrations of gelatin and agar. Synthetic melanin, India ink, absorbing dyes, and Intralipid were added to provide optical absorption and scattering in the skin layers. Bovine serum albumin was used to increase acoustic attenuation, and 40 µm diameter silica microspheres were used to induce acoustic backscatter. Phantom vessels consisting of thin-walled polydimethylsiloxane tubing were embedded at depths of 2-6 mm beneath the skin, and blood mimicking fluid was passed through the vessels. The phantoms were characterized through uniaxial compression and tension experiments, rheological frequency sweep studies, diffuse reflectance spectroscopy, and ultrasonic pulse-echo measurements. Results were then compared to in vivo and ex vivo literature data. RESULTS: The elastic and dynamic shear behavior of the phantom skin layers and vessel wall closely approximated the behavior of porcine skin tissues and human vessels. Similarly, the optical properties of the phantom tissue components in the wavelength range of 400-1100 nm, as well as the acoustic properties in the frequency range of 2-9 MHz, were comparable to human tissue data. Normalized root mean square percent errors between the phantom results and the literature reference values ranged from 1.06% to 9.82%, which for many measurements were less than the sample variability. Finally, the mechanical and imaging characteristics of the phantoms were found to remain stable after 30 days of storage at 21 °C. CONCLUSIONS: The phantoms described in this work simulate the mechanical, optical, and acoustic properties of human skin tissues, vessel tissue, and blood. In this way, the phantoms are uniquely suited to serve as test models for multimodal imaging techniques and image-guided interventions.

Metabolic Flux Distribution during Defatting of Steatotic Human Hepatoma (HepG2) Cells.

Methods that rapidly decrease fat in steatotic hepatocytes may be helpful to recover severely fatty livers for transplantation. Defatting kinetics are highly dependent upon the extracellular medium composition; however, the pathways involved are poorly understood. Steatosis was induced in human hepatoma cells (HepG2) by exposure to high levels of free fatty acids, followed by defatting using plain medium containing no fatty acids, or medium supplemented with a cocktail of defatting agents previously described before. We measured the levels of 28 extracellular metabolites and intracellular triglyceride, and fed the data into a steady-state mass balance model to estimate strictly intracellular fluxes. We found that during defatting, triglyceride content decreased, while beta-oxidation, the tricarboxylic acid cycle, and the urea cycle increased. These fluxes were augmented by defatting agents, and even more so by hyperoxic conditions. In all defatting conditions, the rate of extracellular glucose uptake/release was very small compared to the internal supply from glycogenolysis, and glycolysis remained highly active. Thus, in steatotic HepG2 cells, glycolysis and fatty acid oxidation may co-exist. Together, these pathways generate reducing equivalents that are supplied to mitochondrial oxidative phosphorylation.

Urban Dissolved Silica: Quantifying the Role of Groundwater and Runoff in Wastewater Influent.

Human impacts on silicon (Si) cycling are just being explored. In particular, we know little about the role of urban environments in altering the flux of Si from land to sea. Here we describe the annual load of dissolved Si (DSi) in the influent of the second largest wastewater treatment plant (by volume) in the United States (Deer Island Wastewater Facility, Boston, MA). We partition the ∼69 500 kmol DSi year(-1) influent load between three sources: runoff (12%), groundwater infiltration (39%), and sewage (49%). Based on these results, we hypothesized that instead of being delivered to local rivers, DSi in groundwater and runoff is redirected to the combined stormwater-sewage overflow system. To test this hypothesis we compared long-term (2007-2012) observations of DSi flux from the three urban rivers surrounding Boston to modeled DSi fluxes based on land use and land cover. As predicted, the modeled fluxes were higher than the measured fluxes indicating that the sewage infrastructure of Boston diverts watershed DSi to the treatment plant. This research increases our understanding of human changes to the Si cycle, demonstrates the potential usefulness of DSi as a groundwater infiltration tracer within sewage treatment systems, and highlights the underappreciated interannual variability of riverine DSi fluxes.

Differential Leukocyte Counting via Fluorescent Detection and Image Processing on a Centrifugal Microfluidic Platform.

Centrifugal microfluidics has received much attention in the last decade for the automation of blood testing at the point-of-care, specifically for the detection of chemistries, proteins, and nucleic acids. However, the detection of common blood cells on-disc, particularly leukocytes, remains a challenge. In this paper, we present two analytical methods for enumerating leukocytes on a centrifugal platform using a custom-built fluorescent microscope, acridine orange nuclear staining, and image processing techniques. In the first method, cell analysis is performed in glass capillary tubes; in the second, acrylic chips are used. A bulk-cell analysis approach is implemented in both cases where the pixel areas of fractionated lymphocyte/monocyte and granulocyte layers are correlated with cell counts. Generating standard curves using porcine blood sample controls, we observed strong linear fits to measured cell counts using both methods. Analyzing the pixel intensities of the fluorescing white cell region, we are able to differentiate lymphocytes from monocytes via pixel clustering, demonstrating the capacity to perform a 3-part differential. Finally, a discussion of pros and cons of the bulk-cell analysis approach concludes the paper.

The System Design and Evaluation of a 7-DOF Image-Guided Venipuncture Robot.

Accessing the venous bloodstream to deliver fluids or obtain a blood sample is the most common clinical routine practiced in the U.S. Practitioners continue to rely on manual venipuncture techniques, but success rates are heavily dependent on clinician skill and patient physiology. In the U.S., failure rates can be as high as 50% in difficult patients, making venipuncture the leading cause of medical injury. To improve the rate of first-stick success, we have developed a portable autonomous venipuncture device that robotically servos a needle into a suitable vein under image guidance. The device operates in real time, combining near-infrared and ultra-sound imaging, image analysis, and a 7-degree-of-freedom (DOF) robotic system to perform the venipuncture. The robot consists of a 3-DOF gantry to image the patient's peripheral forearm veins and a miniaturized 4-DOF serial arm to guide the cannula into the selected vein under closed-loop control. In this paper, we present the system architecture of the robot and evaluate the accuracy and precision through tracking, free-space positioning, and in vitro phantom cannulation experiments. The results demonstrate sub-millimeter accuracy throughout the operating workspace of the manipulator and a high rate of success when cannulating phantom veins in a skin-mimicking tissue model.

Real-time Needle Steering in Response to Rolling Vein Deformation by a 9-DOF Image-Guided Autonomous Venipuncture Robot.

Venipuncture is the most common invasive medical procedure performed in the United States and the number one cause of hospital injury. Failure rates are particularly high in pediatric and elderly patients, whose veins tend to deform, move, or roll as the needle is introduced. To improve venipuncture accuracy in challenging patient populations, we have developed a portable device that autonomously servos a needle into a suitable vein under image guidance. The device operates in real time, combining near-infrared and ultrasound imaging, computer vision software, and a 9 degrees-of-freedom robot that servos the needle. In this paper, we present the kinematic and mechanical design of the latest generation robot. We then investigate in silico and in vitro the mechanics of vessel rolling and deformation in response to needle insertions performed by the robot. Finally, we demonstrate how the robot can make real-time adjustments under ultrasound image guidance to compensate for subtle vessel motions during venipuncture.

Elevated sensitivity of macrosteatotic hepatocytes to hypoxia/reoxygenation stress is reversed by a novel defatting protocol.

Macrosteatotic livers exhibit elevated intrahepatic triglyceride (TG) levels in the form of large lipid droplets (LDs), reduced adenosine triphosphate (ATP) levels, and elevated reactive oxygen species (ROS) levels, and this contributes to their elevated sensitivity to ischemia/reperfusion injury during transplantation. Reducing macrosteatosis in living donors through dieting has been shown to improve transplant outcomes. Accomplishing the same feat for deceased donor grafts would require ex vivo exposure to potent defatting agents. Here we used a rat hepatocyte culture system exhibiting a macrosteatotic LD morphology, elevated TG levels, and an elevated sensitivity to hypoxia/reoxygenation (H/R) to test for such agents and ameliorate H/R sensitivity. Macrosteatotic hepatocyte preconditioning for 48 hours with a defatting cocktail that was previously developed to promote TG catabolism reduced the number of macrosteatotic LDs and intracellular TG levels by 82% and 27%, respectively, but it did not ameliorate sensitivity to H/R. Supplementation of this cocktail with l-carnitine, together with hyperoxic exposure, yielded a similar reduction in the number of macrosteatotic LDs and a 57% reduction in intrahepatic TG storage, likely by increasing the supply of acetyl coenzyme A to mitochondria, as indicated by a 70% increase in ketone body secretion. Furthermore, this treatment reduced ROS levels by 32%, increased ATP levels by 27% (to levels near those of lean controls), and completely abolished H/R sensitivity as indicated by approximately 85% viability after H/R and the reduction of cytosolic lactate dehydrogenase release to levels seen in lean controls. Cultures maintained for 48 hours after H/R were approximately 83% viable and exhibited superior urea secretion and bile canalicular transport in comparison with untreated macrosteatotic cultures. In conclusion, these findings show that the elevated sensitivity of macrosteatotic hepatocytes to H/R can be overcome by defatting agents, and they suggest a possible route for the recovery of discarded macrosteatotic grafts.

Effect of added alkalizer and surfactant on dissolution and absorption of the potassium salt of a weakly basic poorly water-soluble drug.

Telcagepant potassium salt (MK-0974) is an oral calcitonin gene-related peptide receptor inhibitor investigated for the treatment of acute migraine. Under gastric pH conditions, the salt rapidly gels, then converts to an insoluble neutral form that creates an impervious shell on the tablet surface, resulting in a slow and variable release dissolution rate and poor bioavailability. Early attempts to develop a solid dosage form, including solid dispersion and nanosuspension formulations, resulted in low exposures in preclinical studies. Thus, a liquid-filled soft gelatin capsule (SGC) formulation (oblong 20) was used for clinical studies. However, a solid dosage form was desirable for commercialization. The slow dissolution of the tablet formulations was overcome by using a basifying agent, arginine, and inclusion of a nonionic surfactant, poloxamer 407. The combination of arginine and poloxamer in the formulation created a local transient basic microenvironment that promoted the dissolution of the salt and prevented rapid precipitation of the neutral form on the tablet surface to form the gel layer. The tablet formulation achieved fast absorption and comparable exposure to the SGC formulation. The final optimized 280 mg tablet formulation was successfully demonstrated to be bioequivalent to the 300 mg SGC formulation.