RNA sequencing of avian paramyxovirus (Paramyxoviridae, Avulavirinae) isolates from wild mallards in Belgium, 2021: complete genomes and coinfections.

We used untargeted RNA sequencing to characterize three Avulavirinae isolates from pooled samples obtained from wild mallards in Belgium in 2021. The complete genome sequences of two avian Orthoavulavirus-1 (AOAV-1) strains and one avian Paraavulavirus-4 (APMV-4) strain were determined confirming hemagglutination inhibition testing of the virus isolates. In addition, the applied sequencing strategy identified an avian influenza virus (AIV) coinfection in all three virus isolates, confirming weak-positive AIV realtime RT-PCR results from the original sample material. In one AOAV-1 isolate, partial sequences covering all genome segments of an AIV of subtype H11N9 could be de novo assembled from the sequencing data. Besides an AIV coinfection, RNA metagenomic data from the APMV-4 isolate also showed evidence of Alpharetrovirus and Megrivirus coinfection. In total, two AOAV-1 of Class II, genotype I.2 and one APMV-4 complete genome sequences were assembled and compared to publicly available sequences, highlighting the importance of surveillance for poultry pathogens in wild birds. Beyond the insights from full genome characterization of virus isolates, untargeted RNA sequencing strategies provide additional insights in the RNA virome of clinical samples as well as their derived virus isolates that are particularly useful when targeting wild avifauna reservoirs of poultry pathogens.

Efficacy of three different cervical dislocation methods for on-farm killing of layer chicks.

Unfit chicks with low viability are often euthanized in the layer industry. An effective euthanasia protocol is characterized by rapid, irreversible insensibility, followed by prompt death. This study was conducted to evaluate the efficacy of three cervical dislocation methods for killing layer chicks (2-3-day-old, avg BW ± SD; 44 ± 3 g, n = 40): manual cervical dislocation (CD), assisted manual cervical dislocation (ACD; the bird's ventral neck is placed on a blunt table edge and the back of the neck pressed firmly), and mechanical cervical dislocation by Koechner Euthanizing Device (KED-model-S). All three killing methods were assessed on anesthetized chicks (intramuscular injections of medetomidine [0.3 mg/kg BW] and ketamine [30 mg/kg BW] were used to induce clinical anesthesia). CD and ACD were also evaluated using conscious chicks to compare the killing methods and to determine the effect of anesthesia on response variables. There were no differences in time to loss of pupillary light reflex, cessation of heartbeat, or duration of gasping between conscious chicks killed with CD and ACD, but these values were all longer for conscious compared to anesthetized chicks. KED resulted in longer latencies to loss of pupillary light reflex, cessation of heartbeat, and duration of gasping. Radiographs revealed that both CD and ACD resulted in cervical luxation, mainly below the C4 vertebra, whereas KED did not cause luxation in any of the 8 chicks tested. Chicks killed by CD and ACD presented more subdural hemorrhage (SDH) at the site of cervical dislocation than those killed by KED. None of the killing methods resulted in brain trauma. Compared to CD and ACD, KED resulted in longer latency to brain death and less anatomical pathology indicating a lower efficacy of KED as an on-farm killing method.

Frequency division multiplexing readout of a transition edge sensor bolometer array with microstrip-type electrical bias lines.

We demonstrate multiplexed readout of 43 transition edge sensor (TES) bolometers operating at 90 mK using a frequency division multiplexing (FDM) readout chain with bias frequencies ranging from 1 to 3.5 MHz and a typical frequency spacing of 32 kHz. We improve the previously reported performance of our FDM system by two important steps. First, we replace the coplanar wires with microstrip wires, which minimize the cross talk from mutual inductance. From the measured electrical cross talk (ECT) map, the ECT of all pixels is carrier leakage dominated. Only five pixels show an ECT level higher than 1%. Second, we reduce the thermal response speed of the TES detectors by a factor of 20 by increasing the heat capacity of the TES, which allows us to bias all TES detectors below 50% in transition without oscillations. We compare the current-voltage curves and noise spectra of the TESs measured in single-pixel mode and multiplexing mode. We also compare the noise equivalent power (NEP) and the saturation power of the bolometers in both modes, where 38 pixels show less than 10% difference in NEP and 5% difference in saturation power when measured in the two different modes. The measured noise spectrum is in good agreement with the simulated noise based on measured parameters from an impedance measurement, confirming that our TES is dominated by phonon noise.

Influences of the COVID-19 Pandemic on Intuitive Exercise and Physical Activity among College Students.

The emergence of the COVID-19 pandemic has significantly impacted the health behaviors of people around the world, including their physical activity patterns. Intuitive exercise, a facet of one's relationship with physical activity, is defined as one's awareness, mindset (positive versus negative), and mindfulness when engaged in movement. The study's purpose was to explore whether self-reported physical activity and psychological mindsets around exercise changed during the pandemic. College students (n = 216) described their relationship with exercise before and during the pandemic through anonymous completion of the Intuitive Exercise Scale (IEXS) and open-ended questions to provide in-depth contextualized responses about exercise habits. Participants reported significantly higher scores on intuitive exercise during the pandemic, such as on the Body Trust subscale (M = 3.43), compared to pre-pandemic levels (M = 3.20), p < 0.001. Moreover, varied themes related to physical activity were uncovered such as exercising for fun, exercise influenced by emotion, and loss of motivation to exercise. Important takeaways of the study include the diversity of responses to the pandemic (i.e., some participants reported an increase in physical activity levels and more positive exercise attitudes while others experienced the opposite), the need to promote self-care, and the need for positive coping strategies.

Impact of Bowel Urgency on Quality of Life and Clinical Outcomes in Patients With Ulcerative Colitis.

Background: Bowel urgency is commonly experienced by patients with ulcerative colitis (UC) and is associated with reduced health-related quality of life (QoL). Mirikizumab, a humanized monoclonal antibody directed against the p19 subunit of IL-23, significantly reduced bowel urgency in a double-blind, randomized, placebo-controlled Phase 2 clinical trial in patients with moderate-to-severe UC (NCT02589665). Methods: All patients (N = 249) reported symptoms including absence or presence of bowel urgency. Absence of urgency was defined as no urgency for the 3 consecutive days prior to each scheduled visit. Missing urgency data were imputed as present. After 12 weeks of induction treatment, patients who achieved clinical response continued maintenance mirikizumab treatment through Week 52. We assessed the relationship of urgency with QoL, clinical outcomes, and inflammatory biomarkers at Weeks 12 and 52. Results: Patients with absence of urgency demonstrated significantly greater improvement in Inflammatory Bowel Disease Questionnaire (IBDQ) scores even after adjusting for rectal bleeding (RB) and stool frequency (SF), significantly higher rates of all clinical outcomes at Weeks 12 and 52, and a greater decrease in inflammatory biomarkers C-reactive protein and fecal calprotectin compared to those with presence of urgency. Absence of urgency at Week 12 was associated with improved IBDQ scores at Week 52, while Week 12 RB or SF status was not. Conclusions: Absence of urgency is strongly associated with improvement in QoL as well as clinical measures of UC disease activity. These findings suggest urgency may be a useful surrogate marker of disease activity and an important treatment target for UC.

Frequency shift algorithm: Application to a frequency-domain multiplexing readout of x-ray transition-edge sensor microcalorimeters.

In the frequency-domain multiplexing (FDM) scheme, transition-edge sensors (TESs) are individually coupled to superconducting LC filters and AC biased at MHz frequencies through a common readout line. To make efficient use of the available readout bandwidth and to minimize the effect of non-linearities, the LC resonators are usually designed to be on a regular grid. The lithographic processes, however, pose a limit on the accuracy of the effective filter resonance frequencies. Off-resonance bias carriers could be used to suppress the impact of intermodulation distortions, which, nonetheless, would significantly affect the effective bias circuit and the detector spectral performance. In this paper, we present a frequency shift algorithm (FSA) to allow off-resonance readout of TESs, while preserving the on-resonance bias circuit and spectral performance, demonstrating its application to the FDM readout of an x-ray TES microcalorimeter array. We discuss the benefits in terms of mitigation of the impact of intermodulation distortions at the cost of increased bias voltage and the scalability of the algorithm to multi-pixel FDM readout. We show that with FSA, in the multi-pixel and frequencies shifted on-grid, the line noises due to intermodulation distortion are placed away from the sensitive region in the TES response and the x-ray performance is consistent with the single-pixel, on-resonance level.

Electrical cross talk of a frequency division multiplexing readout for a transition edge sensor bolometer array.

We have characterized and mapped the electrical cross talk (ECT) of a frequency division multiplexing (FDM) system with a transition edge sensor (TES) bolometer array, which is intended for space applications. By adding a small modulation at 120 Hz to the AC bias voltage of one bolometer and measuring the cross talk response in the current noise spectra of the others simultaneously, we have for the first time mapped the ECT level of 61 pixels with a nominal frequency spacing of 32 kHz in a 61 × 61 matrix and a carrier frequency ranging from 1 MHz to 4 MHz. We find that about 94% of the pixels show an ECT level of less than 0.4%. Only the adjacent pixels reach this level, and the ECT for the rest of the pixels is less than 0.1%. We also observe higher ECT levels, up to 10%, between some of the pixels, which have bundled long, parallel coplanar wires connecting TES bolometers to inductor-capacitor filters. In this case, the high mutual inductances dominate. To mitigate this source of ECT, the coplanar wires should be replaced by microstrip wires in the array. Our study suggests that an FDM system can have a relatively low ECT level, e.g., around 0.4% if the frequency spacing is 30 kHz. Our results successfully demonstrate a low electrical cross talk for a space FDM technology.

Assessment of empiric body mass index-based thromboprophylactic dosing of enoxaparin after bariatric surgery: evidence for dosage adjustment using anti-factor Xa in high-risk patients.

BACKGROUND: Despite thromboprophylaxis, postoperative deep vein thrombosis and pulmonary embolism occur after bariatric surgery, perhaps because of failure to achieve optimal prophylactic levels in the obese population. OBJECTIVES: The aim of this study was to evaluate the adequacy of prophylactic dosing of enoxaparin in patients with severe obesity by performing an antifactor Xa (AFXa) assay. SETTING: An academic medical center METHODS: In this observational study, all bariatric surgery cases at an academic center between December 2016 and April 2017 who empirically received prophylactic enoxaparin (adjusted by body mass index [BMI] threshold of 50 kg/m2) were studied. The AFXa was measured 3-5 hours after the second dose of enoxaparin. RESULTS: A total of 105 patients were included; 85% were female with a median age of 47 years. In total, 16 patients (15.2%) had AFXa levels outside the prophylactic range: 4 (3.8%) cases were in the subprophylactic and 12 (11.4%) cases were in the supraprophylactic range. Seventy patients had a BMI <50 kg/m2 and empirically received enoxaparin 40 mg every 12 hours; AFXa was subprophylactic in 4 (5.7%) and supraprophylactic in 6 (8.6%) of these patients. Of the 35 patients with a BMI ≥50 who empirically received enoxaparin 60 mg q12h, no AFXa was subprophylactic and 6 (17.1%) were supraprophylactic. Five patients (4.8%) had major bleeding complications. One patient developed pulmonary embolism on postoperative day 35. CONCLUSION: BMI-based thromboprophylactic dosing of enoxaparin after bariatric surgery could be suboptimal in 15% of patients with obesity. Overdosing of prophylactic enoxaparin can occur more commonly than underdosing. AFXa testing can be a practical way to measure adequacy of pharmacologic thromboprophylaxis, especially in patients who are at higher risk for venous thromboembolism or bleeding.

Schedule feasibility and workflow for additive manufacturing of titanium plates for ranioplasty in canine skull tumors.

BACKGROUND: Additive manufacturing has allowed for the creation of a patient-specific custom solution that can resolve many of the limitations previously reported for canine cranioplasty. The purpose of this pilot study was to determine the schedule feasibility and workflow in manufacturing patient-specific titanium implants for canines undergoing cranioplasty immediately following craniectomy. RESULTS: Computed tomography scans from patients with tumors of the skull were considered and 3 cases were selected. Images were imported into a DICOM image processing software and tumor margins were determined based on agreement between a board-certified veterinary radiologist and veterinary surgical oncologist. Virtual surgical planning was performed and a bone safety margin was selected. A defect was created to simulate the planned intraoperative defect. Stereolithography format files of the skulls were then imported into a plate design software. In collaboration with a medical solution centre, a custom titanium plate was designed with the input of an applications engineer and veterinary surgery oncologist. Plates were printed in titanium and post-processed at the solution centre. Total planning time was approximately 2 h with a manufacturing time of 2 weeks. CONCLUSIONS: Based on the findings of this study, with access to an advanced 3D metal printing medical solution centre that can provide advanced software and printing, patient-specific additive manufactured titanium implants can be planned, created, processed, shipped and sterilized for patient use within a 3-week turnaround.

Genomic monitoring to understand the emergence and spread of Usutu virus in the Netherlands, 2016-2018.

Usutu virus (USUV) is a mosquito-borne flavivirus circulating in Western Europe that causes die-offs of mainly common blackbirds (Turdus merula). In the Netherlands, USUV was first detected in 2016, when it was identified as the likely cause of an outbreak in birds. In this study, dead blackbirds were collected, screened for the presence of USUV and submitted to Nanopore-based sequencing. Genomic sequences of 112 USUV were obtained and phylogenetic analysis showed that most viruses identified belonged to the USUV Africa 3 lineage, and molecular clock analysis evaluated their most recent common ancestor to 10 to 4 years before first detection of USUV in the Netherlands. USUV Europe 3 lineage, commonly found in Germany, was less frequently detected. This analyses further suggest some extent of circulation of USUV between the Netherlands, Germany and Belgium, as well as likely overwintering of USUV in the Netherlands.

Reliability and usefulness of upper gastro intestinal contrast studies to assess pouch size in patients with weight loss failure after Roux-en-Y gastric bypass.

Background: Weight loss failure or weight regain occurs in up to 25% of patients with a Roux-en-Y gastric bypass (RYGB). Post-operative anatomical changes, like pouch or stoma dilatation, might contribute. Aim of this study is to assess reliability and usefulness of upper gastro intestinal (UGI) contrast studies to detect pouch dilatation.Methods: Retrospective case-control study of patients with weight loss failure between 2010 and 2015 (failure group, n = 101) and a control group (n = 101) with adequate weight loss. Pouch dilatation was systematically reassessed. Clinical parameters were extracted from the electronic patient records.Results: Systematic reassessment showed 23/101 (23%) pouch dilatation in the failure group, compared to 11/101 (11%) in the control group (p = .024). Revision surgery was performed in 43/101 patients in the failure group. After this surgery, only 8% of patients with pouch dilatation achieved adequate weight loss, whereas 39% of patients without pouch dilatation achieved adequate weight loss (p = .07). There was no difference in return to adequate weight loss between patients treated surgically and conservatively (30% vs 28%).Conclusion: Systematic reassessment of UGI contrast studies showed 23% pouch dilatation in patients with weight loss failure after RYGB. However, low interobserver agreement and discrepancy in success rate of revision surgery greatly questions the reliability and usefulness of this diagnostic modality.

[Internet use after prostate cancer : Search for information and trust in disease-related information in long-term survivors]. / Internetnutzung nach Prostatakrebs : Informationssuche und Vertrauen in erkrankungsrelevante Informationen bei Langzeitüberlebenden.

BACKGROUND AND OBJECTIVE: The internet provides numerous sources of information about prostate cancer (PCa). The present study investigated internet use among long-term PCa survivors, trust in online PCa-related information, and associated factors. MATERIALS AND METHODS: Based on the German national research project Familial Prostate Cancer long-term PCa survivors were asked about their internet use in 2017. Associations with sociodemographic (age at survey, children, intimate relationship, education) and disease-related parameters (time since diagnosis, PCa family history, progress) were analyzed using multivariable logistic regression. RESULTS: In all, 4636 long-term PCa survivors were included in the analysis (mean age 76.9 years; standard deviation 6.6 years). Mean follow-up was 14.0 years. Of long-term PCa survivors, 62.1% were using the internet. Among non-users 23.5% expressed strong concerns, among users only 2.8%. Furthermore, 47.2% of internet users sought information about PCa, 18.0% of them indicated difficulties while searching for information. More than half of the users found the online information inappropriate. Lower age, shorter time since diagnosis, progress, and a more frequent internet use were associated with search for information. Only one-third fully trusted online information. Trust in online information was associated with high age, higher educational level, and frequent search for online information. Many survivors stressed that they were primarily trusting their treating urologist. CONCLUSIONS: Two-thirds of long-term PCa survivors are using the internet. A significant proportion expressed difficulties finding proper and reliable information. Urologists should be familiar with online resources on PCa in order to offer advice to patients and to recommend adequate information on the internet.

Towards high quality real-time whole genome sequencing during outbreaks using Usutu virus as example.

Recently, protocols for amplicon based whole genome sequencing using Nanopore technology have been described for Ebola virus, Zika virus, yellow fever virus and West Nile virus. However, there is some debate regarding reliability of sequencing using this technology, which is important for applications beyond diagnosis such as linking lineages to outbreaks, tracking transmission pathways and pockets of circulation, or mapping specific markers. To our knowledge, no in depth analyses of the required read coverage to compensate for the error profile in Nanopore sequencing have been described. Here, we describe the validation of a protocol for whole genome sequencing of USUV using Nanopore sequencing by direct comparison to Illumina sequencing. To that point we selected brain tissue samples with high viral loads, typical for birds which died from USUV infection. We conclude that the low-cost MinION Nanopore sequencing platform can be used for characterization and tracking of Usutu virus outbreaks.

LiGAPS-Beef, a mechanistic model to explore potential and feed-limited beef production 3: model evaluation.

LiGAPS-Beef (Livestock simulator for Generic analysis of Animal Production Systems - Beef cattle) is a generic, mechanistic model designed to quantify potential and feed-limited growth, which provides insight in the biophysical scope to increase beef production (i.e. yield gap). Furthermore, it enables identification of the bio-physical factors that define and limit growth, which provides insight in management strategies to mitigate yield gaps. The aim of this paper, third in a series of three, is to evaluate the performance of LiGAPS-Beef with independent experimental data. After model calibration, independent data were used from six experiments in Australia, one in Uruguay and one in the Netherlands. Experiments represented three cattle breeds, and a wide range of climates, feeding strategies and cattle growth rates. The mean difference between simulated and measured average daily gains (ADGs) was 137 g/day across all experiments, which equals 20.1% of the measured ADGs. The root mean square error was 170 g/day, which equals 25.0% of the measured ADGs. LiGAPS-Beef successfully simulated the factors that defined and limited growth during the experiments on a daily basis (genotype, heat stress, digestion capacity, energy deficiency and protein deficiency). The simulated factors complied well to the reported occurrence of heat stress, energy deficiency and protein deficiency at specific periods during the experiments. We conclude that the level of accuracy of LiGAPS-Beef is acceptable, and provides a good basis for acquiring insight in the potential and feed-limited production of cattle in different beef production systems across the world. Furthermore, its capacity to identify factors that define or limit growth and production provides scope to use the model for yield gap analysis.

LiGAPS-Beef, a mechanistic model to explore potential and feed-limited beef production 2: sensitivity analysis and evaluation of sub-models.

The model LiGAPS-Beef (Livestock simulator for Generic analysis of Animal Production Systems - Beef cattle) has been developed to assess potential and feed-limited growth and production of beef cattle in different areas of the world and to identify the processes responsible for the yield gap. Sensitivity analysis and evaluation of model results with experimental data are important steps after model development. The first aim of this paper, therefore, is to identify which parameters affect the output of LiGAPS-Beef most by conducting sensitivity analyses. The second aim is to evaluate the accuracy of the thermoregulation sub-model and the feed intake and digestion sub-model with experimental data. Sensitivity analysis was conducted using a one-at-a-time approach. The upper critical temperature (UCT) simulated with the thermoregulation sub-model was most affected by the body core temperature and parameters affecting latent heat release from the skin. The lower critical temperature (LCT) and UCT were considerably affected by weather variables, especially ambient temperature and wind speed. Sensitivity analysis for the feed intake and digestion sub-model showed that the digested protein per kg feed intake was affected to a larger extent than the metabolisable energy (ME) content. Sensitivity analysis for LiGAPS-Beef was conducted for ¾ Brahman×» Shorthorn cattle in Australia and Hereford cattle in Uruguay. Body core temperature, conversion of digestible energy to ME, net energy requirements for maintenance, and several parameters associated with heat release affected feed efficiency at the herd level most. Sensitivity analyses have contributed, therefore, to insight which parameters are to be investigated in more detail when applying LiGAPS-Beef. Model evaluation was conducted by comparing model simulations with independent data from experiments. Measured heat production in experiments corresponded fairly well to the heat production simulated with the thermoregulation sub-model. Measured ME contents from two data sets corresponded well to the ME contents simulated with the feed intake and digestion sub-model. The relative mean absolute errors were 9.3% and 6.4% of the measured ME contents for the two data sets. In conclusion, model evaluation indicates the thermoregulation sub-model can deal with a wide range of weather conditions, and the feed intake and digestion sub-model with a variety of feeds, which corresponds to the aim of LiGAPS-Beef to simulate cattle in different beef production systems across the world.

LiGAPS-Beef, a mechanistic model to explore potential and feed-limited beef production 1: model description and illustration.

The expected increase in the global demand for livestock products calls for insight in the scope to increase actual production levels across the world. This insight can be obtained by using theoretical concepts of production ecology. These concepts distinguish three production levels for livestock: potential (i.e. theoretical maximum) production, which is defined by genotype and climate only; feed-limited production, which is limited by feed quantity and quality; and actual production. The difference between the potential or limited production and the actual production is the yield gap. The objective of this paper, the first in a series of three, is to present a mechanistic, dynamic model simulating potential and feed-limited production for beef cattle, which can be used to assess yield gaps. A novelty of this model, named LiGAPS-Beef (Livestock simulator for Generic analysis of Animal Production Systems - Beef cattle), is the identification of the defining factors (genotype and climate) and limiting factors (feed quality and available feed quantity) for cattle growth by integrating sub-models on thermoregulation, feed intake and digestion, and energy and protein utilisation. Growth of beef cattle is simulated at the animal and herd level. The model is designed to be applicable to different beef production systems across the world. Main model inputs are breed-specific parameters, daily weather data, information about housing, and data on feed quality and quantity. Main model outputs are live weight gain, feed intake and feed efficiency (FE) at the animal and herd level. Here, the model is presented, and its use is illustrated for Charolais and Brahman × Shorthorn cattle in France and Australia. Potential and feed-limited production were assessed successfully, and we show that FE of herds is highest for breeds most adapted to the local climate conditions. LiGAPS-Beef also identified the factors that define and limit growth and production of cattle. Hence, we argue the model has scope to be used as a tool for the assessment and analysis of yield gaps in beef production systems.

Early changes in N-terminal pro-B-type natriuretic peptide levels after transcatheter aortic valve replacement and its impact on long-term mortality.

BACKGROUND: N-terminal pro-hormone B-type natriuretic peptide (NT-proBNP) levels correlate with higher peri-procedural mortality after transcatheter aortic valve replacement (TAVR). The long-term prognostic value of NT-proBNP within the first days after TAVR, however, remains unclear. This study examined early changes in NT-proBNP prior to and within 6 days after TAVR, the diagnostic value of this biomarker regarding aortic regurgitation (AR), and its prognostic value regarding one-year mortality. METHODS AND RESULTS: NT-proBNP concentrations were measured in 504 consecutive patients undergoing transapical (TA) or transfemoral (TF) TAVR before and directly after TAVR as well as 4 h and 1, 2, 3, and 6 days after TAVR. The follow-up period was 1 year. NT-proBNP was elevated in all patients at baseline (median 2141 ng/L [IQR 1021-5319 ng/L]). NT-proBNP changes in the first 6 days after TAVR showed significant differences depending on the approach, with a greater and more prolonged rise evident in TA-TAVR patients. NT-proBNP was an independent predictor of mortality in TA patients with AR, with an AUC of 0.794 (95% CI 0.663-0.925; P = 0.003) when measured on day 3 after TAVR. For TF patients with AR and reduced left ventricular systolic function, the AUC for prediction of mortality was 0.897 (95% CI 0.778-1.0; P = 0.004) on day 2. CONCLUSIONS: The prognostic information of early post-procedural NT-proBNP concentrations is superior to pre-procedural values regarding all-cause mortality within 1 year. Post-procedural NT-proBNP must be interpreted in relation to the TAVR approach. NT-proBNP predicts mortality in TF-TAVR patients with AR and reduced left ventricular function.

Metformin does not reduce inflammation in diabetics with abdominal aortic aneurysm or at high risk of abdominal aortic aneurysm formation.

INTRODUCTION: The protective effect of diabetes mellitus on abdominal aortic aneurysm formation and growth has been repeatedly observed in population studies but continues to be poorly understood. However, recent investigations have suggested that metformin, a staple antihyperglycemic medication, may be independently protective against abdominal aortic aneurysm formation and growth. Therefore, we describe the effect of metformin in abdominal aortic aneurysm and at-risk patients on markers of inflammation, the driver of early abdominal aortic aneurysm formation and growth. METHODS: Peripheral blood was collected from patients previously diagnosed with abdominal aortic aneurysm or presenting for their U.S. Preventive Task Force-recommended abdominal aortic aneurysm screening. Plasma and circulating peripheral blood mononuclear cells were isolated using Ficoll density centrifugation. Circulating plasma inflammatory and regulatory cytokines were assessed with enzyme-linked immunosorbent assays. CD4+ cell phenotyping was performed using flow cytometric analysis and expressed as a proportion of total CD4+ cells. To determine the circulating antibody to self-antigen response, a modified enzyme-linked immunosorbent assay was performed against antibodies to collagen type V and elastin fragments. RESULTS: Peripheral blood was isolated from 266 patients without diabetes mellitus ( n=182), with diabetes mellitus not treated with metformin ( n=34), and with diabetes mellitus actively taking metformin ( n=50) from 2015 to 2017. We found no differences in the expression of Tr1, Th17, and Treg CD4+ fractions within diabetics ± metformin. When comparing inflammatory cytokines, we detected no differences in IL-1ß, IL-6, IL-17, IL-23, IFN-γ, and TNF-α. Conversely, no differences were observed pertaining to the expression to regulatory cytokines IL-4, IL-10, IL-13, TSG-6, or TGF-ß. Lastly, no differences in expression of collagen type V and elastin fragment antigen and/or antibodies were detected with metformin use in diabetics. CONCLUSION: Metformin in diabetics at-risk for abdominal aortic aneurysm or diagnosed with abdominal aortic aneurysm does not seem to alter the peripheral inflammatory environment.

Description of human AAA by cytokine and immune cell aberrations compared to risk-factor matched controls.

BACKGROUND: The pathogenesis driving the formation of abdominal aortic aneurysms continues to be poorly understood. Therefore, we systemically define the cytokine and circulating immune cell environment observed in human abdominal aortic aneurysm compared with risk-factor matched controls. METHODS: From 2015 to 2017, a total of 274 patients donated blood to the Indiana University Center for Aortic Disease. Absolute concentrations of circulating cytokines were determined, using enzyme-linked immunosorbent assays while the expression of circulating immune cell phenotypes were assayed via flow cytometric analysis. RESULTS: Human abdominal aortic aneurysm is characterized by a significant depletion of the antigen-specific, CD4+ Tr1 regulatory lymphocyte that corresponds to an upregulation of the antigen-specific, inflammatory Th17 cell. We found no differences in the incidence of Treg, B10, and myeloid-derived suppressor regulatory cells. Similarly, no disparities were noted in the following inflammatory cytokines: IL-1ß, C-reactive protein, tumor necrosis factor α, interferon γ, and IL-23. However, significant upregulation of the inflammatory cytokines osteopontin, IL-6, and IL-17 were noted. Additionally, no changes were observed in the regulatory cytokines IL-2, IL-4, IL-13, TNF-stimulated gene 6 protein, and prostaglandin E2, but we did observe a significant decrease in the essential regulatory cytokine IL-10. CONCLUSION: In this investigation, we systematically characterize the abdominal aortic aneurysm-immune environment and present preliminary evidence that faulty immune regulation may also contribute to aneurysm formation and growth.

Development and growth trends in angiotensin II-induced murine dissecting abdominal aortic aneurysms.

Abdominal aortic aneurysms are pathological dilations that can suddenly rupture, causing more than 15,000 deaths in the U.S. annually. Current treatment focuses on observation until an aneurysm's size warrants surgical intervention. Thus, there is a need for therapeutic intervention to inhibit growth of smaller aneurysms. An experimental aneurysm model that infuses angiotensin II into apolipoprotein E-deficient mice is widely used to investigate underlying pathological mechanisms and potential therapeutics, but this model has two caveats: (1) aneurysms do not always form, and (2) aneurysm severity and growth is inconsistent among animals. Here we use high-frequency ultrasound to collect data from angiotensin II-induced aneurysms to develop prediction models of both aneurysm formation and growth. Baseline measurements of aortic diameter, volume/length, and strain were used with animal mass and age in a quadratic discriminant analysis and logistic regression to build two statistical models to predict disease status. Longitudinal ultrasound data were also acquired from mice with aneurysms to quantify aneurysm diameter, circumferential strain, blood flow velocity, aneurysm volume/length, and thrombus and open-false lumen volumes over 28 days. Measurements taken at aneurysm diagnosis were used with branching artery information to produce a multiple linear regression model to predict final aneurysm volume/length. All three statistical models could be useful in future aneurysm therapeutic studies to better delineate the effects of preventative and suppressive treatments from normal variations in the angiotensin II aneurysm model.