Novices' learning curve in single-port surgery using three surgical training programs.

BACKGROUND: The objective of this study was to evaluate the novices' learning curves and proficiency level reached in laparoendoscopic single-site (LESS) surgery using three surgical training programs. MATERIAL AND METHODS: Participants were randomly divided into three groups, who trained in a specific practice regimen for 12 days using a laparoscopic box simulator and three tasks. Group A trained in three stages using conventional laparoscopic surgery (CLS) with straight instruments, and LESS with straight and articulating instruments for four days each. Group B trained in two stages in LESS with straight and articulating instruments for six days each. Group C trained only in LESS with articulating instruments exclusively for all 12 days. Performance was registered daily during the 12 days to evaluate the participants' progress. RESULTS: Pre- and post-training analysis of the three groups showed significant differences in performance, denoting the significant improvement in their LESS skills, with no difference between the groups. Group C reached a high level of technical competence with their specific training program in LESS, obtaining a lower asymptote and slow learning rate. CONCLUSION: Specific training programs in LESS settings using articulated instruments showed a slower learning rate than the other programs but better proficiency in the technique with the best surgical performance.

Methodology for fast development of digital solutions in integrated continuous downstream processing.

The methodology for production of biologics is going through a paradigm shift from batch-wise operation to continuous production. Lot of efforts are focused on integration, intensification, and continuous operation for decreased foot-print, material, equipment, and increased productivity and product quality. These integrated continuous processes with on-line analytics become complex processes, which requires automation, monitoring, and control of the operation, even unmanned or remote, which means bioprocesses with high level of automation or even autonomous capabilities. The development of these digital solutions becomes an important part of the process development and needs to be assessed early in the development chain. This work discusses a platform that allows fast development, advanced studies, and validation of digital solutions for integrated continuous downstream processes. It uses an open, flexible, and extendable real-time supervisory controller, called Orbit, developed in Python. Orbit makes it possible to communicate with a set of different physical setups and on the same time perform real-time execution. Integrated continuous processing often implies parallel operation of several setups and network of Orbit controllers makes it possible to synchronize complex process system. Data handling, storage, and analysis are important properties for handling heterogeneous and asynchronous data generated in complex downstream systems. Digital twin applications, such as advanced model-based and plant-wide monitoring and control, are exemplified using computational extensions in Orbit, exploiting data and models. Examples of novel digital solutions in integrated downstream processes are automatic operation parameter optimization, Kalman filter monitoring, and model-based batch-to-batch control.

Objective Evaluation of Laparoscopic Experience Based on Muscle Electromyography and Accelerometry Performing Circular Pattern Cutting Tasks: A Pilot Study.

Purpose.The aim of this work is to present a new physical laparoscopy simulator with an electromyography (EMG)/accelerometry-based muscle activity recording system, EvalLap EMG-ACC, and perform objective evaluation of laparoscopic skills based on the quantification of muscle activity of participants with different levels of laparoscopic experience. Methods. EMG and ACC signals were obtained from 14 participants (6 experts, 8 medical students) performing circular pattern cutting tasks using a laparoscopic box trainer with the Trigno (Delsys Inc, Natick, MA) portable wireless system of 16 wireless sensors. Sensors were placed on the proximal and distal muscles of the upper extremities. Seven evaluation metrics were proposed and compared between skilled and novice surgeons. Results. The proximal and distal arm muscles (trapezius, deltoids, biceps, and forearms) were most active while executing laparoscopic tasks. Laparoscopic experience was associated with differences in EMG amplitude (Aavg), muscle activity (iEMG), hand acceleration (iACH), user movement (iAC), and muscle fatigue. For the cutting task, the deltoid, bicep, forearm EMG amplitude, and user movement significantly differed between experience groups. Conclusion. This pilot study demonstrates that different muscle groups are preferentially activated during laparoscopic tasks depending on the level of surgical experience. Expert surgeons showed less muscle activity compared with novices. EvalLap EMG-ACC represents a promising means to distinguish surgeons with basic cutting skills from those who have not yet developed these skills.

Steerable Surgical Instrument for Conventional and Single-Site Minimally Invasive Surgery.

Background. This article aims to present an innovative design of a steerable surgical instrument for conventional and single-site minimally invasive surgery (MIS), which improves the dexterity and maneuverability of the surgeon while offering a solution to the limitations of current tools. Methods. The steerable MIS instrument consists of a deflection structure with a curved sliding joints design that articulates the distal tip in two additional degrees of freedom (DoFs), relative to the instrument shaft, using transmission by cables. A passive ball-joint mechanism articulates the handle relative to the instrument shaft, improves wrist posture, and prevents collision of instrument handles during single-site MIS procedures. The two additional DoFs of the articulating tip are activated by a thumb-controlled device, using a joystick design mounted on the handle. This steerable MIS instrument was developed by additive manufacturing in a 3D printer using PLA polymer. Results. Prototype testing showed a maximum tip deflection of 60° in the left and right directions, with a total deflection of 120°. With the passive ball-joint fully offset, the steerable tip achieved a deflection of 90° for the right and 40° for the left direction, with a total deflection of 130°. Furthermore, the passive ball-joint mechanism in the handle obtained a maximum range of motion of 60°. Conclusions. This steerable MIS instrument concept offers an alternative to enhance the application fields of conventional and single-site MIS, increasing manual dexterity of the surgeon and the ability to reach narrow anatomies from other directions.

Dengue in Western Uganda: a prospective cohort of children presenting with undifferentiated febrile illness.

BACKGROUND: The spatial distribution and burden of dengue in sub-Saharan Africa remains highly uncertain, despite high levels of ecological suitability. The goal of this study was to describe the epidemiology of dengue among a cohort of febrile children presenting to outpatient facilities located in areas of western Uganda with differing levels of urbanicity and malaria transmission intensity. METHODS: Eligible children were first screened for malaria using rapid diagnostic tests. Children with a negative malaria result were tested for dengue using a combination NS1/IgM/IgG rapid test (SD Bioline Dengue Duo). Confirmatory testing by RT-PCR was performed in a subset of participants. Antigen-capture ELISA was performed to estimate seroprevalence. RESULTS: Only 6 of 1416 (0.42%) children had a positive dengue rapid test, while none of the RT-PCR results were positive. ELISA testing demonstrated reactive IgG antibodies in 28 (2.2%) participants with the highest prevalence seen at the urban site in Mbarara (19 of 392, 4.9%, p < 0.001). CONCLUSIONS: Overall, these findings suggest that dengue, while present, is an uncommon cause of non-malarial, pediatric febrile illness in western Uganda. Further investigation into the eocological factors that sustain low-level transmission in urban settings are urgently needed to reduce the risk of epidemics.

Glucocorticoid-induced CREB activation and myostatin expression in C2C12 myotubes involves phosphodiesterase-3/4 signaling.

Muscle atrophy in metabolic conditions like chronic kidney disease (CKD) and diabetes are associated with glucocorticoid production, dysfunctional insulin/Akt/FoxO3 signaling and increased myostatin expression. We recently found that CREB, a transcription factor proposed to regulate myostatin expression, is highly phosphorylated in some wasting conditions. Based on a novel Akt-PDE3/4 signaling paradigm, we hypothesized that reduced Akt signaling contributes to CREB activation and myostatin expression. C2C12 myotubes were incubated with dexamethasone (Dex), an atrophy-inducing synthetic glucocorticoid. Akt/CREB signaling and myostatin expression were evaluated by immunoblot and qPCR analyses. Inhibitors of Akt, phosphodiesterase (PDE)-3/4, and protein kinase A (PKA) signaling were used to test our hypothesis. Incubating myotubes with Dex for 3-24 h inhibited Akt phosphorylation and enhanced CREB phosphorylation as well as myostatin mRNA and protein. Inhibition of PI3K/Akt signaling with LY294002 similarly increased CREB phosphorylation. Isobutyl-methylxanthine (IBMX, a pan PDE inhibitor), milrinone (PDE3 inhibitor) and rolipram (PDE4 inhibitor) augmented CREB phosphorylation and myostatin expression. Inhibition of protein kinase A by PKI reverted Dex- or IBMX-induced CREB phosphorylation and myostatin expression. Our study provides evidence supporting a newly identified mechanism by which a glucocorticoid-related reduction in Akt signaling contributes to myostatin expression via CREB activation.

Optimization of a biotechnological multiproduct batch plant design for the manufacture of four different products: A real case scenario.

In this work a biotechnological multiproduct batch plant that manufactures four different recombinant proteins for human application is described in some detail. This batch plant design is then optimized with regards to the size of equipment using a mixed-integer linear programming (MILP) formulation recently developed by us in order to find a hypothetical new biotechnological batch plant based on the information of real known processes for the production of the four recombinant protein products. The real plant was divided for practical purposes into two sub-processes or facilities: a fermentation facility and a purification facility. Knowing the specific steps conforming the downstream processing of each product, size, and time factors were computed and used as parameters to solve the aforementioned MILP reformulation. New constraints were included to permit the selection of some equipment-such as centrifuges and membrane filters-in a discrete set of sizes. For equipment that can be built according to customer needs-such as reactors-the original formulation was retained. Computational results show the ability of this optimization methodology to deal with real data giving reliable solutions for a multi-product batch plant composed of 44 unit operations in a relatively small amount of time showing that in the case studied it is possible to save up to a 66% of the capital investment in equipment given the cost data used. Biotechnol. Bioeng. 2017;114: 1252-1263. © 2017 Wiley Periodicals, Inc.

Clinicopathological Spectrum of Mitral Valve Myxoma.

Cardiac myxomas are the most common primary tumors of the heart. Although these tumors have been reported in all cardiac chambers, myxomas arising from heart valves are extremely rare. Here, the details are described of a patient with mitral valve myxoma, and a review is provided of 99 cases reported in the literature. Mitral valve myxomas most often occur in middle-aged patients, with a slight female predominance. Most of the tumors arise in an isolated fashion on the atrial side of the anterior mitral valve leaflet. Embolic events were the most frequent manifestation, followed by obstructive symptoms. Unlike atrial wall myxomas, these tumors have a lower incidence of constitutional manifestations. Tumor resection with valve preservation is the usual method of surgical treatment in isolated tumors, while most patients with multiple valve myxomas underwent valve replacement. Mitral valve myxomas should be considered in the differential diagnosis of mitral valve tumors, with an awareness of a relatively high risk of embolic events. This location should not raise suspicion for a heritable myxoma syndrome. Prompt surgical resection is warranted to reduce complications.

Glucose uptake and lipid metabolism are impaired in epicardial adipose tissue from heart failure patients with or without diabetes.

Type 2 diabetes mellitus is a complex metabolic disease, and cardiovascular disease is a leading complication of diabetes. Epicardial adipose tissue surrounding the heart displays biochemical, thermogenic, and cardioprotective properties. However, the metabolic cross-talk between epicardial fat and the myocardium is largely unknown. This study sought to understand epicardial adipose tissue metabolism from heart failure patients with or without diabetes. We aimed to unravel possible differences in glucose and lipid metabolism between human epicardial and subcutaneous adipocytes and elucidate the potential underlying mechanisms involved in heart failure. Insulin-stimulated [(14)C]glucose uptake and isoproterenol-stimulated lipolysis were measured in isolated epicardial and subcutaneous adipocytes. The expression of genes involved in glucose and lipid metabolism was analyzed by reverse transcription-polymerase chain reaction in adipocytes. In addition, epicardial and subcutaneous fatty acid composition was analyzed by high-resolution proton nuclear magnetic resonance spectroscopy. The difference between basal and insulin conditions in glucose uptake was significantly decreased (P= 0.006) in epicardial compared with subcutaneous adipocytes. Moreover, a significant (P< 0.001) decrease in the isoproterenol-stimulated lipolysis was also observed when the two fat depots were compared, and it was strongly correlated with lipolysis, lipid storage, and inflammation-related gene expression. Moreover, the fatty acid composition of these tissues was significantly altered by diabetes. These results emphasize potential metabolic differences between both fat depots in the presence of heart failure and highlight epicardial fat as a possible therapeutic target in situ in the cardiac microenvironment.

Pediatric inguinal hernia repair with a single-incision approach using an Endo Close™ suturing device.

BACKGROUND: Diverse techniques have been described for pediatric inguinal hernia repair, based on extraperitoneal [1-4] and intraperitoneal [5-8] methodologies. In this video, we describe a novel technique to repair pediatric inguinal hernia using an Endo Close™ suturing device by percutaneous puncture with a single incision. METHODS: With a transumbilical approach, a 5-mm trocar is inserted for a 30° laparoscope. A 3-mm incision is made, and the Endo Close™ suturing device (Covidien, Minneapolis, MN, USA), with a 2-0 polypropylene suture retained by the stylet, is inserted perpendicularly to the skin. An extraperitoneal dissection is made on a side the inguinal ring and the needle of the device penetrates the peritoneum through the inferior border. Then, the stylet mechanism is pushed to free the lasso inside the cavity. At the same incision site, the needle of the Endo Close™ is inserted again, but an extraperitoneal dissection is made on the other side of the ring, ensuring that the needle penetrates at the same exit orifice. Now, the suture lasso is recovered and retracted to close the ring. Finally, the suture is extracted and knots are tied extracorporeally at the level of the skin. RESULTS: A total of 34 patients (20 females and 14 males) underwent surgery with this procedure. Operative time for unilateral repair was 10-15 and 25-30 min for the bilateral repair (29 unilateral/5 bilateral). The patients experienced minimal postoperative pain. The follow-up period was 12 months with no complications, no recurrence and without cases of postoperative hydrocele. There were no injuries to the structures as vessels or vas deferens, and the esthetic outcome was excellent. CONCLUSIONS: The technique presents a simple, safe and reliable method to repair inguinal hernias in children. The long-term results of this novel technique will be evaluated in future studies.

New role of bone morphogenetic protein 7 in brown adipogenesis and energy expenditure.

Adipose tissue is central to the regulation of energy balance. Two functionally different types of fat are present in mammals: white adipose tissue, the primary site of triglyceride storage, and brown adipose tissue, which is specialized in energy expenditure and can counteract obesity. Factors that specify the developmental fate and function of white and brown adipose tissue remain poorly understood. Here we demonstrate that whereas some members of the family of bone morphogenetic proteins (BMPs) support white adipocyte differentiation, BMP7 singularly promotes differentiation of brown preadipocytes even in the absence of the normally required hormonal induction cocktail. BMP7 activates a full program of brown adipogenesis including induction of early regulators of brown fat fate PRDM16 (PR-domain-containing 16; ref. 4) and PGC-1alpha (peroxisome proliferator-activated receptor-gamma (PPARgamma) coactivator-1alpha; ref. 5), increased expression of the brown-fat-defining marker uncoupling protein 1 (UCP1) and adipogenic transcription factors PPARgamma and CCAAT/enhancer-binding proteins (C/EBPs), and induction of mitochondrial biogenesis via p38 mitogen-activated protein (MAP) kinase-(also known as Mapk14) and PGC-1-dependent pathways. Moreover, BMP7 triggers commitment of mesenchymal progenitor cells to a brown adipocyte lineage, and implantation of these cells into nude mice results in development of adipose tissue containing mostly brown adipocytes. Bmp7 knockout embryos show a marked paucity of brown fat and an almost complete absence of UCP1. Adenoviral-mediated expression of BMP7 in mice results in a significant increase in brown, but not white, fat mass and leads to an increase in energy expenditure and a reduction in weight gain. These data reveal an important role of BMP7 in promoting brown adipocyte differentiation and thermogenesis in vivo and in vitro, and provide a potential new therapeutic approach for the treatment of obesity.

Real-time monitoring of gradient chromatography using dual Kalman-filters.

Real-time state estimation in chromatography is a useful tool to improve monitoring of biopharmaceutical downstream processes, combining mechanistic model predictions with real-time data acquisition to obtain an estimation that surpasses that of either approach individually. One common technique for real-time state estimation is Kalman filtering. However, non-linear adsorption isotherms pose a significant challenge to Kalman filters, which are dependent on fast algorithm execution to function. In this work, we apply Kalman filtering of non-constant elution conditions using a non-linear adsorption isotherm using a novel approach where dual Kalman filters are used to estimate the states of the adsorption modifier, salt, and the components to be separated. We performed offline tuning of the Kalman filters on real chromatogram data from a linear gradient, ion-exchange separation of two proteins. The tuning was then validated by running the Kalman filters in parallel with a chromatographic separation in real time. The resulting, tuned, dual Kalman filters improved the L2 norm by 53 % over the open-loop model prediction, when compared to the true elution profiles. The Kalman filters were also applicable in real-time with a signal sampling frequency of 5 s, enabling accurate and robust estimation and paving the way for future applications beyond monitoring, such as real-time optimal pooling control.

Automatic procedure for modelling, calibration, and optimization of a three-component chromatographic separation.

The current landscape of biopharmaceutical production necessitates an ever-growing set of tools to meet the demands for shorter development times and lower production costs. One path towards meeting these demands is the implementation of digital tools in the development stages. Mathematical modelling of process chromatography, one of the key unit operations in the biopharmaceutical downstream process, is one such tool. However, obtaining parameter values for such models is a time-consuming task that grows in complexity with the number of compounds in the mixture being purified. In this study, we tackle this issue by developing an automated model calibration procedure for purification of a multi-component mixture by linear gradient ion exchange chromatography. The procedure was implemented using the Orbit software (Lund University, Department of Chemical Engineering), which both generates a mathematical model structure and performs the experiments necessary to obtain data for model calibration. The procedure was extended to suggest operating points for the purification of one of the components in the mixture by means of multi-objective optimization using three different objectives. The procedure was tested on a three-component protein mixture and was able to generate a calibrated model capable of reproducing the experimental chromatograms to a satisfactory degree, using a total of six assays. An additional seventh experiment was performed to validate the model response under one of the suggested optimum conditions, respecting a 95 % purity requirement. All of the above was automated and set in motion by the push of a button. With these results, we have taken a step towards fully automating model calibration and thus accelerating digitalization in the development stages of new biopharmaceuticals.

Spatial distribution of Triatoma dimidiata peridomestic colonies modulated by distance between susceptible microhabitat patches.

Triatoma dimidiata is a vector of the hemoparasite Trypanosoma cruzi, the causal agent of Chagas disease. It settles reproductive colonies in the peridomicile of the premises. The peridomicile is comprised of a random set of artificial and natural features that overlap and assemble a network of microenvironmental suitable sites (patches) that interact with each other and favor the structure and proliferation of T. dimidiata colonies. The heterogeneity of patch characteristics hinders the understanding and identification of sites susceptible to colonization. In this study, a classification system using a random forest algorithm was used to identify peridomiciles susceptible to colonization to describe the spatial distribution of these sites and their relationship with the colonies of T. dimidiata in ten localities of Yucatan. From 1,000 peridomiciles reviewed, the classification showed that 13.9 % (139) of the patches were highly susceptible (HSP), and 86.1 % (861) were less susceptible (LSP). All localities had at least one HSP. The occupancy by patch type showed that the percentage of total occupancy and by colonies was higher in the HSP, while the occupancy by adult T. dimidiata without evidence of nymphs or exuviae (propagules) was higher in the LSP. A generalized additive model (GAM) revealed that the percentage of occupied patches increases as the abundance of individuals in the localities increases however, the percentage of occupied patches in LSP is lower than occupied in HSP. Distance analyses revealed that colonies and propagules were located significantly closer (approximately 200 m) to a colony in a HSP than any colony in a LSP. The distribution of T. dimidiata in the localities was defined by the distribution of patch type; as the occupancy in these patches increased, a network of peridomestic populations was configured, which may be promoted by a greater abundance of insects inside the localities. These results reveal that the spatial distribution of T. dimidiata individuals and colonies in the peridomicile at the locality scale corresponds to a metapopulation pattern within the localities through a system of patches mediated by distance and level of the vectors' occupancy.

Image Guided Interpedicular Screw Placement Simulation System for Training and Skill Evaluation. Proof of Concept.

BACKGROUND: The SpineST-01 system is an image-guided vertebrae cannulation training system. During task execution, the computer calculates performance-based metrics displaying different visual perspectives (lateral view, axial view, anteroposterior view) with the position of the instrument inside the vertebra. Finally, a report with the metrics is generated as performance feedback. METHODS: A training box holds a 3D printed spine section. The computer works with 2 orthogonally disposed cameras, tracking passive markers placed on the instrument. Eight metrics were proposed to evaluate the execution of the surgical task. A preliminary study with 25 participants divided into 3 groups (12 novices, 10 intermediates, and 3 expert) was conducted to determine the feasibility of the system and to evaluate and assess the performance differences of each group using Kruskal-Wallis analysis and Mann-Whitney U analysis. In both analyses, a P value ≤ 0.05 was considered statistically significant. RESULTS: When comparing experts versus novices and all 3 groups, statistical analysis showed significant differences in 6 of the 8 metrics: axial angle error (°), lateral angle error (°), average speed (mm/second), progress between shots (mm), Time (seconds), and shots. The metrics that did not show any statistically significant difference were time between shots (seconds), and speed between shots (mm/second). Also, the average result comparison placed the experts as the best performance group. CONCLUSIONS: Initial testing of the SpineST-01 demonstrated potential for the system to practice image-guided cannulation tasks on lumbar vertebrae. Results showed objective differences between experts, intermediates, and novices in the proposed metrics, making this system a feasible option for developing basic navigation system skills without the risk of radiation exposure and objectively evaluating task performance.

Evidence of Ongoing Transmission of Zika Virus in Mérida, Mexico.

Since the Zika virus (ZIKV) pandemic in 2015-2017, there has been a near absence of reported cases in the Americas outside of Brazil. However, the conditions for Aedes-borne transmission persist in Latin America, and the threat of ZIKV transmission is increasing as population immunity wanes. Mexico has reported only 70 cases of laboratory-confirmed ZIKV infection since 2020, with no cases recorded in the Yucatán peninsula. Here, we provide evidence of active ZIKV transmission, despite the absence of official case reports, in the city of Mérida, Mexico, the capital of the state of Yucatán. Capitalizing on an existing cohort, we detected cases in participants with symptoms consistent with flavivirus infection from 2021 to 2022. Serum samples from suspected cases were tested for ZIKV RNA by polymerase chain reaction or ZIKV-reactive IgM by ELISA. To provide more specific evidence of exposure, focus reduction neutralization tests were performed on ELISA-positive samples. Overall, we observed 25 suspected ZIKV infections for an estimated incidence of 2.8 symptomatic cases per 1,000 persons per year. Our findings emphasize the continuing threat of ZIKV transmission in the setting of decreased surveillance and reporting.

Bone morphogenetic protein 7 (BMP7) reverses obesity and regulates appetite through a central mTOR pathway.

Body weight is regulated by coordinating energy intake and energy expenditure. Transforming growth factor ß (TGFß)/bone morphogenetic protein (BMP) signaling has been shown to regulate energy balance in lower organisms, but whether a similar pathway exists in mammals is unknown. We have previously demonstrated that BMP7 can regulate brown adipogenesis and energy expenditure. In the current study, we have uncovered a novel role for BMP7 in appetite regulation. Systemic treatment of diet-induced obese mice with BMP7 resulted in increased energy expenditure and decreased food intake, leading to a significant reduction in body weight and improvement of metabolic syndrome. Similar degrees of weight loss with reduced appetite were also observed in BMP7-treated ob/ob mice, suggesting a leptin-independent mechanism utilized by BMP7. Intracerebroventricular administration of BMP7 to mice led to an acute decrease in food intake, which was mediated, at least in part, by a central rapamycin-sensitive mTOR-p70S6 kinase pathway. Together, these results underscore the importance of BMP7 in regulating both food intake and energy expenditure, and suggest new therapeutic approaches for obesity and its comorbidities.

Automated quality analysis in continuous downstream processes for small-scale applications.

Development of integrated, continuous biomanufacturing (ICB) processes brings along the challenge of streamlining the acquisition of data that can be used for process monitoring, product quality testing and process control. Manually performing sample acquisition, preparation, and analysis during process and product development on ICB platforms requires time and labor that diverts attention from the development itself. It also introduces variability in terms of human error in the handling of samples. To address this, a platform for automatic sampling, sample preparation and analysis for use in small-scale biopharmaceutical downstream processes was developed. The automatic quality analysis system (QAS) consisted of an ÄKTA Explorer chromatography system for sample retrieval, storage, and preparation, as well as an Agilent 1260 Infinity II analytical HPLC system for analysis. The ÄKTA Explorer system was fitted with a superloop in which samples could be stored, conditioned, and diluted before being sent to the injection loop of the Agilent system. The Python-based software Orbit, developed at the department of chemical engineering at Lund university, was used to control and create a communication framework for the systems. To demonstrate the QAS in action, a continuous capture chromatography process utilizing periodic counter-current chromatography was set up on an ÄKTA Pure chromatography system to purify the clarified harvest from a bioreactor for monoclonal antibody production. The QAS was connected to the process to collect two types of samples: 1) the bioreactor supernatant and 2) the product pool from the capture chromatography. Once collected, the samples were conditioned and diluted in the superloop before being sent to the Agilent system, where both aggregate content and charge variant composition were determined using size-exclusion and ion-exchange chromatography, respectively. The QAS was successfully implemented during a continuous run of the capture process, enabling the acquisition of process data with consistent quality and without human intervention, clearing the path for automated process monitoring and data-based control.

Incidental eagle carcass detection can contribute to fatality estimation at operating wind energy facilities.

Risk of birds colliding with wind turbines, especially protected species like bald eagle and golden eagle in the U.S., is a fundamental wildlife challenge the wind industry faces when developing and operating projects. The U.S. Fish and Wildlife Service requires wind energy facilities that obtain eagle take permits document permit compliance through fatality monitoring. If trained Operations and Maintenance (O&M) staff can reliably detect and report carcasses during their normal routines, and their detection probability can be estimated, then their 'incidental detections' could contribute substantially towards demonstrating permit compliance. Our primary objective was to quantify incidental detection of eagle carcasses by O&M staff under a variety of landscape contexts and environmental conditions throughout a single year. We used the incidental detection probabilities, along with raptor carcass persistence data and area adjustments, to calculate overall probability of incidental detection (i.e., incidental g). We used feathered decoys as eagle-carcass surrogates for monthly detection trials at 6 study sites throughout the U.S. We evaluated the primary drivers of incidental detection using logit regression models including season, viewshed complexity, and a derived variable called the "density quartile" as covariates. We used an Evidence of Absence-based approach to estimate the overall probability of incidental detection. The incidental detection probabilities ranged from 0.28 to 0.78 (mean = 0.48). Detection probabilities decreased as viewshed complexity increased and as distance from the turbine increased. The resulting overall probability of incidental detection ranged from 0.07 to 0.47 (mean = 0.31). The primary drivers of variability in incidental g were detection probability and the area adjustment. Results of our research show that O&M staff were effective at detecting trial carcasses incidentally. Incorporating incidental detection in eagle fatality monitoring efforts is a reliable means of improving estimates of a facility's direct impacts on eagles.

Game bird carcasses are less persistent than raptor carcasses, but can predict raptor persistence dynamics.

Researchers conduct post-construction fatality monitoring (PCFM) to determine a wind energy facility's direct impacts on wildlife. Results of PCFM can be used to evaluate compliance with permitted take, potentially triggering adaptive management measures or offsetting mitigation; reducing uncertainty in fatality rates benefits wind companies, wildlife agencies, and other stakeholders. As part of PCFM, investigators conduct carcass persistence trials to account for imperfect detection during carcass surveys. In most PCFM studies, pen-raised game birds and other non-raptor surrogates have been used to estimate persistence of all large birds, including raptors. However, there is a growing body of evidence showing carcass persistence varies by bird type; raptor fatality estimates based on game bird carcass persistence may therefore be biased high. We conducted raptor and game bird carcass persistence field trials for 1 year at 6 wind energy facilities. Raptor carcass persistence varied by habitat and season, whereas the best-supported game bird model only included habitat. Raptor persistence probabilities were higher than corresponding game bird persistence probabilities for 13 of the 16 habitat and season combinations. Analysis of a curated large bird persistence meta-dataset showed that raptor carcass persistence varied by season, habitat, and region. The probability of persisting through a 30-day search interval ranged from 0.44 to 0.99 for raptors and from 0.16 to 0.79 for game birds. Raptor persistence was significantly higher than game bird persistence for 95% of the sampled strata. We used these carcass persistence estimates to develop linear mixed-effects models that predict raptor persistence probabilities based on estimated game bird persistence probabilities. Our scaling model provides an important statistical method to address gaps in raptor persistence data at sites in a broad range of landscape contexts in the continental United States and should be used to inform fatality estimation when site-specific raptor persistence data are limited or absent.